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  • Long ssDNA for knockins
  • Knockin screening kit
  • Genome-wide sgRNA library system
  • Recombinant Cas9 protein
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  • Cas9 antibodies
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Home › Products › Gene function › Gene editing › CRISPR-Cas9 › Lentiviral systems

Gene editing

  • CRISPR-Cas9
    • Long ssDNA for knockins
    • Knockin screening kit
    • Genome-wide sgRNA library system
    • Recombinant Cas9 protein
    • GMP recombinant Cas9
    • Mutation detection kits
    • In vitro transcription and screening kits
    • Cas9-sgRNA gesicle production
    • Cas9 antibodies
    • Plasmid systems
    • Lentiviral systems
    • AAV systems
    • RNA transfection
    • Genotype confirmation kit
    • Indel identification kit
  • Cre recombinase
    • AAV2-Cre recombinase
    • Cre Recombinase Gesicles
    • Cre recombinase RLPs
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Lentiviral systems

Lenti-X and Lenti-X Tet-On 3G CRISPR/Cas9 systems

The Lenti-X CRISPR/Cas9 System and Lenti-X Tet-On 3G CRISPR/Cas9 System are complete systems for lentiviral-mediated CRISPR/Cas9 genome editing. By using lentivirus to deliver sgRNA and Cas9, these systems allow you to achieve targeted genome editing in cell lines that are difficult to transfect. Each system includes Lenti-X Packaging Single Shots for generating high titers of virus and plasmid sets designed to express Cas9 and your custom sgRNA in target cells.

The Lenti-X CRISPR/Cas9 System and Lenti-X Tet-On 3G CRISPR/Cas9 System are complete systems for lentiviral-mediated CRISPR/Cas9 genome editing. By using lentivirus to deliver sgRNA and Cas9, these systems allow you to achieve targeted genome editing in cell lines that are difficult to transfect. Each system includes Lenti-X Packaging Single Shots for generating high titers of virus and plasmid sets designed to express Cas9 and your custom sgRNA in target cells. The plasmid for sgRNA expression, pLVX-hyg-sgRNA1, comes prelinearized for easy insertion of your sgRNA sequence and is supplied in sufficient quantity for the construction of 10 different plasmids, each with its own sgRNA sequence.

Whereas the Lenti-X CRISPR/Cas9 System involves the constitutive expression of Cas9 in target cells, the Lenti-X Tet-On 3G CRISPR/Cas9 System incorporates the Tet-On 3G transactivator protein, which enables users to induce Cas9 expression via the addition of doxycycline. By allowing for tight control of Cas9 expression, the Lenti-X Tet-On 3G CRISPR/Cas9 System enables users to minimize the likelihood of toxicity and off-target effects associated with persistent Cas9 expression in cell culture.

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Cat. # Product Size Price License Quantity Details
632630 pLVX-hyg-sgRNA1 Vector System 10 Rxns USD $340.00

License Statement

ID Number  
272 This product (“Product”) and its use, is the subject of U.S. Patents 8,697,359 and 8,771,945 and pending U.S. Patent applications. The purchase of the Product conveys to the buyer the non-transferable right to use Product(s) purchased from Takara Bio USA, Inc. or its Affiliates, and any progeny, modification or derivative of a Product, or any cell or animal made or modified through use of a Product, or any progeny, modification or derivative of such cell or animal (“Related Material”), solely for research conducted by the buyer in accordance with all of the following requirements. No right is given to use this Product or Related Material for any other purpose, including, but not limited to, use in drugs, in vitro diagnostic purposes, therapeutics, or in humans. (i) The buyer shall not sell or otherwise transfer Products (including without limitation any material that contains a Licensed Product in whole or part) or any Related Material to any other person or entity, or use Products or any Related Material to perform services for the benefit of any other person or entity, (ii) the buyer shall use only the purchased amount of the Products and components of the Products, and shall use any Related Material, only for its internal research and not for (a) the practice, performance or provision of any method, process or service, or (b) the manufacture, sale, use, distribution, disposition or importing of any product, in each case (a) or (b) for consideration, or on any other commercial basis (“Commercial Purpose”), (iii) the buyer shall use Licensed Products and any Related Material in compliance with all applicable laws and regulations, including without limitation applicable human health and animal welfare laws and regulations, and (iv) the buyer shall indemnify, defend and hold harmless MIT, Harvard and The Broad and their current and former trustees, directors, officers, faculty, affiliated investigators, students, employees, and agents and their respective successors, heirs and assigns (“Indemnitees”), against any liability, damage, loss, or expense (including without limitation reasonable attorneys’ fees and expenses) incurred by or imposed upon any of the Indemnitees in connection with any claims, suits, investigations, actions, demands or judgments arising out of or related to the exercise of any rights granted to the buyer, or any breach of the rights granted hereunder by the buyer.
391 LIMITED USE LABEL LICENSE: RESEARCH USE ONLY Notice to Purchaser: This product is the subject to a license granted to Takara Bio USA, Inc. and its Affiliates from Caribou Biosciences, Inc., and this product is transferred to the end-user purchaser (“Purchaser”) subject to a “Limited Use Label License” conveying to the Purchaser a limited, non-transferable right to use the product, solely as provided to Purchaser, together with (i) progeny or derivatives of the product generated by the Purchaser (including but not limited to cells), and (ii) biological material extracted or derived from the product or its corresponding progeny or derivatives (including but not limited to cells) (collectively, the product, and (i) and (ii) are referred to as (“Material”) only to perform internal research for the sole benefit of the Purchaser. The Purchaser cannot sell or otherwise transfer Material to a third party or otherwise use the Material for any Excluded Use. “Excluded Use” means any and all: (a) commercial activity including, but not limited to, any use in manufacturing (including but not limited to cell line development for purposes of bioproduction), product testing, or quality control; (b) preclinical or clinical testing or other activity directed toward the submission of data to the U.S. Food and Drug Administration, or any other regulatory agency in any country or jurisdiction where the active agent in such studies comprises the Material; (c) use to provide a service, information, or data to a third party; (d) use for human or animal therapeutic, diagnostic, or prophylactic purposes or as a product for therapeutics, diagnostics, or prophylaxis; (e) activity in an agricultural field trial or any activity directed toward the submission of data to the U.S. Department of Agriculture or any other agriculture regulatory agency; (f) high throughput screening drug discovery purposes (i.e., the screening of more than 10,000 experiments per day) as well as scale-up production activities for commercialization; (g) modification of human germline, including editing of human embryo genomes (with the sole exception of editing human embryonic stem (ES) cell lines for research purposes) or reproductive cells; (h) self-editing; and/or (i) stimulation of biased inheritance of a particular gene or trait or set of genes or traits (“gene drive”). It is the Purchaser’s responsibility to use the Material in accordance with all applicable laws and regulations. For information on obtaining additional rights, including commercial rights, please contact licensing@cariboubio.com or Caribou Biosciences, Inc., 2929 7th Street, Suite 105, Berkeley, CA 94710 USA, Attn: Licensing.
396 Sigma-Aldrich CRISPR Use License Agreement This Product and its use are the subject of one or more of the following issued patents and patent applications: Australia Patent Nos. 2013355214; 2017204031; and 2018229489; Canada Patent Nos. 2,891,347 and 2,977,152; China Patent No. CN105142669; European Patent Nos. EP 2 928 496 B1; EP 3 138 910 B1, 3 138 911 B1, EP 3 138 912 B1, EP 3 360 964 B1, EP 3 363 902 B1; Israel Patent No. IL238856; Singapore Patent No. 11201503824S; South Korea Patent Nos. 10-1844123 and 10-2006880; and U.S. Patent Application Serial Nos. 15/188,911; 15/188,924; 15/188,927; 15/188,931; and 15/456,204 (the “Patent Rights”). The purchase of this Product conveys to you (the “Buyer”) the NON-TRANSFERABLE right to use the Product for Licensed Research Use (see definition below) subject to the conditions set out in this License Agreement. 1. “Licensed Research Use” means any use for research purposes, except: (i) Buyer may not sell or otherwise transfer the Product (including without limitation any material that contains the Product in whole or part) or any Related Material to any other third party (except that you may transfer the Product, or any Related Material to a bona fide collaborator or contract research organization), or use the Products or any Related Material to perform services for the benefit of any other third party; (ii) Buyer may use only the purchased amount of the Product and components of the Product, and shall use any Related Material, only for your internal research within the Field, and not for any Commercial Purposes; (iii) Buyer shall use the Product and any Related Material in compliance with all applicable laws and regulations, including without limitation applicable human health and animal welfare laws and regulations; and (iv) the Buyer shall indemnify, defend, and hold harmless SIGMA and their current and former directors, officers, employees and agents, and their respective successors, heirs and assigns (the “Indemnities”) against any liability, damage, loss, or expense (including without limitation reasonable attorneys’ fees and expenses) incurred by or imposed upon any of the Indemnitees in connection with any claims, suits, investigations, actions, demands or judgments arising out of or related to the exercise of any rights granted to the Buyer hereunder or any breach of this License Agreement by such Buyer. 2. For purposes of Section 1 above, the following definitions shall apply: “Commercial Purposes” means (a) the practice, performance or provision of any method, process or service, or (b) the manufacture, sale, use, distribution, disposition or importing of any product, in each case (a) or (b) for consideration, or on any other commercial basis. “Field” means use as a research tool for research purposes; provided, however, that notwithstanding the foregoing, the Field shall expressly exclude (a) any in vivo and ex vivo human or clinical use, including, without limitation, any administration into humans or any diagnostic or prognostic use, (b) the creation of transgenic rodent models and/or derivatives thereof (including, but not limited to, rodents’ cells and rodents’ organs) by for-profit entities, (c) any in vivo veterinary or livestock use, or non-research agricultural use, or (d) use as a testing service, therapeutic or diagnostic for humans or animals. “Related Materials” means any progeny, modification or derivative of a Product. 3. Your right to use the Product will terminate immediately if you fail to comply with these terms and conditions. You shall, upon such termination of your rights, destroy all Product, Related Materials, and components thereof in your control, and notify SIGMA of such in writing. For information on purchasing a license to this Product for purposes other than Licensed Research Use, contact your local SIGMA sales representative, or call +1 800-325-3010.

The Lenti-X CRISPR/Cas9 System and Lenti-X Tet-On 3G CRISPR/Cas9 System are complete systems for lentiviral-mediated CRISPR/Cas9 genome editing.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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632630: pLVX-hyg-sgRNA1 Vector System

632630: pLVX-hyg-sgRNA1 Vector System
632631 pLVX-puro-Cas9 Vector 20 uL USD $653.00

License Statement

ID Number  
272 This product (“Product”) and its use, is the subject of U.S. Patents 8,697,359 and 8,771,945 and pending U.S. Patent applications. The purchase of the Product conveys to the buyer the non-transferable right to use Product(s) purchased from Takara Bio USA, Inc. or its Affiliates, and any progeny, modification or derivative of a Product, or any cell or animal made or modified through use of a Product, or any progeny, modification or derivative of such cell or animal (“Related Material”), solely for research conducted by the buyer in accordance with all of the following requirements. No right is given to use this Product or Related Material for any other purpose, including, but not limited to, use in drugs, in vitro diagnostic purposes, therapeutics, or in humans. (i) The buyer shall not sell or otherwise transfer Products (including without limitation any material that contains a Licensed Product in whole or part) or any Related Material to any other person or entity, or use Products or any Related Material to perform services for the benefit of any other person or entity, (ii) the buyer shall use only the purchased amount of the Products and components of the Products, and shall use any Related Material, only for its internal research and not for (a) the practice, performance or provision of any method, process or service, or (b) the manufacture, sale, use, distribution, disposition or importing of any product, in each case (a) or (b) for consideration, or on any other commercial basis (“Commercial Purpose”), (iii) the buyer shall use Licensed Products and any Related Material in compliance with all applicable laws and regulations, including without limitation applicable human health and animal welfare laws and regulations, and (iv) the buyer shall indemnify, defend and hold harmless MIT, Harvard and The Broad and their current and former trustees, directors, officers, faculty, affiliated investigators, students, employees, and agents and their respective successors, heirs and assigns (“Indemnitees”), against any liability, damage, loss, or expense (including without limitation reasonable attorneys’ fees and expenses) incurred by or imposed upon any of the Indemnitees in connection with any claims, suits, investigations, actions, demands or judgments arising out of or related to the exercise of any rights granted to the buyer, or any breach of the rights granted hereunder by the buyer.
391 LIMITED USE LABEL LICENSE: RESEARCH USE ONLY Notice to Purchaser: This product is the subject to a license granted to Takara Bio USA, Inc. and its Affiliates from Caribou Biosciences, Inc., and this product is transferred to the end-user purchaser (“Purchaser”) subject to a “Limited Use Label License” conveying to the Purchaser a limited, non-transferable right to use the product, solely as provided to Purchaser, together with (i) progeny or derivatives of the product generated by the Purchaser (including but not limited to cells), and (ii) biological material extracted or derived from the product or its corresponding progeny or derivatives (including but not limited to cells) (collectively, the product, and (i) and (ii) are referred to as (“Material”) only to perform internal research for the sole benefit of the Purchaser. The Purchaser cannot sell or otherwise transfer Material to a third party or otherwise use the Material for any Excluded Use. “Excluded Use” means any and all: (a) commercial activity including, but not limited to, any use in manufacturing (including but not limited to cell line development for purposes of bioproduction), product testing, or quality control; (b) preclinical or clinical testing or other activity directed toward the submission of data to the U.S. Food and Drug Administration, or any other regulatory agency in any country or jurisdiction where the active agent in such studies comprises the Material; (c) use to provide a service, information, or data to a third party; (d) use for human or animal therapeutic, diagnostic, or prophylactic purposes or as a product for therapeutics, diagnostics, or prophylaxis; (e) activity in an agricultural field trial or any activity directed toward the submission of data to the U.S. Department of Agriculture or any other agriculture regulatory agency; (f) high throughput screening drug discovery purposes (i.e., the screening of more than 10,000 experiments per day) as well as scale-up production activities for commercialization; (g) modification of human germline, including editing of human embryo genomes (with the sole exception of editing human embryonic stem (ES) cell lines for research purposes) or reproductive cells; (h) self-editing; and/or (i) stimulation of biased inheritance of a particular gene or trait or set of genes or traits (“gene drive”). It is the Purchaser’s responsibility to use the Material in accordance with all applicable laws and regulations. For information on obtaining additional rights, including commercial rights, please contact licensing@cariboubio.com or Caribou Biosciences, Inc., 2929 7th Street, Suite 105, Berkeley, CA 94710 USA, Attn: Licensing.
396 Sigma-Aldrich CRISPR Use License Agreement This Product and its use are the subject of one or more of the following issued patents and patent applications: Australia Patent Nos. 2013355214; 2017204031; and 2018229489; Canada Patent Nos. 2,891,347 and 2,977,152; China Patent No. CN105142669; European Patent Nos. EP 2 928 496 B1; EP 3 138 910 B1, 3 138 911 B1, EP 3 138 912 B1, EP 3 360 964 B1, EP 3 363 902 B1; Israel Patent No. IL238856; Singapore Patent No. 11201503824S; South Korea Patent Nos. 10-1844123 and 10-2006880; and U.S. Patent Application Serial Nos. 15/188,911; 15/188,924; 15/188,927; 15/188,931; and 15/456,204 (the “Patent Rights”). The purchase of this Product conveys to you (the “Buyer”) the NON-TRANSFERABLE right to use the Product for Licensed Research Use (see definition below) subject to the conditions set out in this License Agreement. 1. “Licensed Research Use” means any use for research purposes, except: (i) Buyer may not sell or otherwise transfer the Product (including without limitation any material that contains the Product in whole or part) or any Related Material to any other third party (except that you may transfer the Product, or any Related Material to a bona fide collaborator or contract research organization), or use the Products or any Related Material to perform services for the benefit of any other third party; (ii) Buyer may use only the purchased amount of the Product and components of the Product, and shall use any Related Material, only for your internal research within the Field, and not for any Commercial Purposes; (iii) Buyer shall use the Product and any Related Material in compliance with all applicable laws and regulations, including without limitation applicable human health and animal welfare laws and regulations; and (iv) the Buyer shall indemnify, defend, and hold harmless SIGMA and their current and former directors, officers, employees and agents, and their respective successors, heirs and assigns (the “Indemnities”) against any liability, damage, loss, or expense (including without limitation reasonable attorneys’ fees and expenses) incurred by or imposed upon any of the Indemnitees in connection with any claims, suits, investigations, actions, demands or judgments arising out of or related to the exercise of any rights granted to the Buyer hereunder or any breach of this License Agreement by such Buyer. 2. For purposes of Section 1 above, the following definitions shall apply: “Commercial Purposes” means (a) the practice, performance or provision of any method, process or service, or (b) the manufacture, sale, use, distribution, disposition or importing of any product, in each case (a) or (b) for consideration, or on any other commercial basis. “Field” means use as a research tool for research purposes; provided, however, that notwithstanding the foregoing, the Field shall expressly exclude (a) any in vivo and ex vivo human or clinical use, including, without limitation, any administration into humans or any diagnostic or prognostic use, (b) the creation of transgenic rodent models and/or derivatives thereof (including, but not limited to, rodents’ cells and rodents’ organs) by for-profit entities, (c) any in vivo veterinary or livestock use, or non-research agricultural use, or (d) use as a testing service, therapeutic or diagnostic for humans or animals. “Related Materials” means any progeny, modification or derivative of a Product. 3. Your right to use the Product will terminate immediately if you fail to comply with these terms and conditions. You shall, upon such termination of your rights, destroy all Product, Related Materials, and components thereof in your control, and notify SIGMA of such in writing. For information on purchasing a license to this Product for purposes other than Licensed Research Use, contact your local SIGMA sales representative, or call +1 800-325-3010.

pLVX-puro-Cas9 Vector is provided as part of the Lenti-X CRISPR/Cas9 System (Cat. # 632629).  This vector is designed for lentiviral-mediated delivery of Cas9 nuclease expressed from a CMV promoter. The Cas9 is the wild-type nuclease from S. pyogenes. Stable integration of Cas9 can be selected for using puromycin.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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632631: pLVX-puro-Cas9 Vector

632631: pLVX-puro-Cas9 Vector
632633 Lenti-X™ Tet-On® 3G CRISPR/Cas9 System 1 System Inquire for Quotation

License Statement

ID Number  
42 Use of the Tetracycline controllable expression systems (the "Tet Technology") is covered by a series of patents including U.S. Patent # 8383364, # 9181556 , European patents EP # 1954811, #2352833 and corresponding patent claims outside these regions which are proprietary to TET Systems GmbH & Co. KG. Academic research institutions are granted an automatic license with the purchase of this product to use the Tet Technology only for internal, academic research purposes, which license specifically excludes the right to sell, or otherwise transfer, the Tet Technology or its component parts to third parties. Notwithstanding the above, academic and not-for profit research institutions whose research using the Tet Technology is sponsored by for profit organizations, which shall receive ownership to any data and results stemming from the sponsored research, shall need a commercial license agreement from TET Systems in order to use the Tet Technology. In accepting this license, all users acknowledge that the Tet Technology is experimental in nature. TET Systems GmbH & Co. KG makes no warranties, express or implied or of any kind, and hereby disclaims any warranties, representations, or guarantees of any kind as to the Tet Technology, patents, or products. All others are invited to request a license from TET Systems GmbH & Co. KG prior to purchasing these reagents or using them for any purpose. Takara Bio USA, Inc. is required by its licensing agreement to submit a report of all purchasers of the Tet-controllable expression system to TET Systems.

For license information, please contact:
GSF/CEO
TET Systems GmbH & Co. KG,
Im Neuenheimer Feld 582
69120 Heidelberg
Germany
Tel: +49 6221 5880400
Fax: +49 6221 5880404
email: info@tetsystems.com
or use the electronic licensing request form via https://www.tetsystems.com/licensing/
63 Use of this product is covered by one or more of the following U.S. Patent Nos. and corresponding patent claims outside the U.S.: 8,562,966, 8,557,231. This product is intended for research purposes only. It may not be used for (i) any human or veterinary use, including without limitation therapeutic and prophylactic use, (ii) any clinical use, including without limitation diagnostic use, (iii) screening of chemical and/or biological compounds for the identification of pharmaceutically active agents (including but not limited to screening of small molecules), target validation, preclinical testing services, or drug development. Any use of this product for any of the above mentioned purposes requires a license from the Massachusetts Institute of Technology.
259 This Product is protected by one or more patents from the family consisting of: JP6454352 and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing common priority with the same family.
272 This product (“Product”) and its use, is the subject of U.S. Patents 8,697,359 and 8,771,945 and pending U.S. Patent applications. The purchase of the Product conveys to the buyer the non-transferable right to use Product(s) purchased from Takara Bio USA, Inc. or its Affiliates, and any progeny, modification or derivative of a Product, or any cell or animal made or modified through use of a Product, or any progeny, modification or derivative of such cell or animal (“Related Material”), solely for research conducted by the buyer in accordance with all of the following requirements. No right is given to use this Product or Related Material for any other purpose, including, but not limited to, use in drugs, in vitro diagnostic purposes, therapeutics, or in humans. (i) The buyer shall not sell or otherwise transfer Products (including without limitation any material that contains a Licensed Product in whole or part) or any Related Material to any other person or entity, or use Products or any Related Material to perform services for the benefit of any other person or entity, (ii) the buyer shall use only the purchased amount of the Products and components of the Products, and shall use any Related Material, only for its internal research and not for (a) the practice, performance or provision of any method, process or service, or (b) the manufacture, sale, use, distribution, disposition or importing of any product, in each case (a) or (b) for consideration, or on any other commercial basis (“Commercial Purpose”), (iii) the buyer shall use Licensed Products and any Related Material in compliance with all applicable laws and regulations, including without limitation applicable human health and animal welfare laws and regulations, and (iv) the buyer shall indemnify, defend and hold harmless MIT, Harvard and The Broad and their current and former trustees, directors, officers, faculty, affiliated investigators, students, employees, and agents and their respective successors, heirs and assigns (“Indemnitees”), against any liability, damage, loss, or expense (including without limitation reasonable attorneys’ fees and expenses) incurred by or imposed upon any of the Indemnitees in connection with any claims, suits, investigations, actions, demands or judgments arising out of or related to the exercise of any rights granted to the buyer, or any breach of the rights granted hereunder by the buyer.
391 LIMITED USE LABEL LICENSE: RESEARCH USE ONLY Notice to Purchaser: This product is the subject to a license granted to Takara Bio USA, Inc. and its Affiliates from Caribou Biosciences, Inc., and this product is transferred to the end-user purchaser (“Purchaser”) subject to a “Limited Use Label License” conveying to the Purchaser a limited, non-transferable right to use the product, solely as provided to Purchaser, together with (i) progeny or derivatives of the product generated by the Purchaser (including but not limited to cells), and (ii) biological material extracted or derived from the product or its corresponding progeny or derivatives (including but not limited to cells) (collectively, the product, and (i) and (ii) are referred to as (“Material”) only to perform internal research for the sole benefit of the Purchaser. The Purchaser cannot sell or otherwise transfer Material to a third party or otherwise use the Material for any Excluded Use. “Excluded Use” means any and all: (a) commercial activity including, but not limited to, any use in manufacturing (including but not limited to cell line development for purposes of bioproduction), product testing, or quality control; (b) preclinical or clinical testing or other activity directed toward the submission of data to the U.S. Food and Drug Administration, or any other regulatory agency in any country or jurisdiction where the active agent in such studies comprises the Material; (c) use to provide a service, information, or data to a third party; (d) use for human or animal therapeutic, diagnostic, or prophylactic purposes or as a product for therapeutics, diagnostics, or prophylaxis; (e) activity in an agricultural field trial or any activity directed toward the submission of data to the U.S. Department of Agriculture or any other agriculture regulatory agency; (f) high throughput screening drug discovery purposes (i.e., the screening of more than 10,000 experiments per day) as well as scale-up production activities for commercialization; (g) modification of human germline, including editing of human embryo genomes (with the sole exception of editing human embryonic stem (ES) cell lines for research purposes) or reproductive cells; (h) self-editing; and/or (i) stimulation of biased inheritance of a particular gene or trait or set of genes or traits (“gene drive”). It is the Purchaser’s responsibility to use the Material in accordance with all applicable laws and regulations. For information on obtaining additional rights, including commercial rights, please contact licensing@cariboubio.com or Caribou Biosciences, Inc., 2929 7th Street, Suite 105, Berkeley, CA 94710 USA, Attn: Licensing.
396 Sigma-Aldrich CRISPR Use License Agreement This Product and its use are the subject of one or more of the following issued patents and patent applications: Australia Patent Nos. 2013355214; 2017204031; and 2018229489; Canada Patent Nos. 2,891,347 and 2,977,152; China Patent No. CN105142669; European Patent Nos. EP 2 928 496 B1; EP 3 138 910 B1, 3 138 911 B1, EP 3 138 912 B1, EP 3 360 964 B1, EP 3 363 902 B1; Israel Patent No. IL238856; Singapore Patent No. 11201503824S; South Korea Patent Nos. 10-1844123 and 10-2006880; and U.S. Patent Application Serial Nos. 15/188,911; 15/188,924; 15/188,927; 15/188,931; and 15/456,204 (the “Patent Rights”). The purchase of this Product conveys to you (the “Buyer”) the NON-TRANSFERABLE right to use the Product for Licensed Research Use (see definition below) subject to the conditions set out in this License Agreement. 1. “Licensed Research Use” means any use for research purposes, except: (i) Buyer may not sell or otherwise transfer the Product (including without limitation any material that contains the Product in whole or part) or any Related Material to any other third party (except that you may transfer the Product, or any Related Material to a bona fide collaborator or contract research organization), or use the Products or any Related Material to perform services for the benefit of any other third party; (ii) Buyer may use only the purchased amount of the Product and components of the Product, and shall use any Related Material, only for your internal research within the Field, and not for any Commercial Purposes; (iii) Buyer shall use the Product and any Related Material in compliance with all applicable laws and regulations, including without limitation applicable human health and animal welfare laws and regulations; and (iv) the Buyer shall indemnify, defend, and hold harmless SIGMA and their current and former directors, officers, employees and agents, and their respective successors, heirs and assigns (the “Indemnities”) against any liability, damage, loss, or expense (including without limitation reasonable attorneys’ fees and expenses) incurred by or imposed upon any of the Indemnitees in connection with any claims, suits, investigations, actions, demands or judgments arising out of or related to the exercise of any rights granted to the Buyer hereunder or any breach of this License Agreement by such Buyer. 2. For purposes of Section 1 above, the following definitions shall apply: “Commercial Purposes” means (a) the practice, performance or provision of any method, process or service, or (b) the manufacture, sale, use, distribution, disposition or importing of any product, in each case (a) or (b) for consideration, or on any other commercial basis. “Field” means use as a research tool for research purposes; provided, however, that notwithstanding the foregoing, the Field shall expressly exclude (a) any in vivo and ex vivo human or clinical use, including, without limitation, any administration into humans or any diagnostic or prognostic use, (b) the creation of transgenic rodent models and/or derivatives thereof (including, but not limited to, rodents’ cells and rodents’ organs) by for-profit entities, (c) any in vivo veterinary or livestock use, or non-research agricultural use, or (d) use as a testing service, therapeutic or diagnostic for humans or animals. “Related Materials” means any progeny, modification or derivative of a Product. 3. Your right to use the Product will terminate immediately if you fail to comply with these terms and conditions. You shall, upon such termination of your rights, destroy all Product, Related Materials, and components thereof in your control, and notify SIGMA of such in writing. For information on purchasing a license to this Product for purposes other than Licensed Research Use, contact your local SIGMA sales representative, or call +1 800-325-3010.
*

The Lenti-X Tet-On 3G CRISPR/Cas9 System is designed for Tet-inducible Cas9 expression along with expression of a custom sgRNA. Because Cas9 is only expressed in the presence of doxycycline, genome editing is achieved on demand and without the side effects of persistent Cas9 expression in cell culture. This kit contains all of the components necessary for generating three types of lentivirus: one for constitutive expression of the Tet-On 3G transactivator protein, a second for Tet-inducible Cas9 expression under the PTRE3GV promoter, and a third for constitutive sgRNA expression under the U6 promoter

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Inducible knockout of CD81 in HEK293 cells

Inducible knockout of CD81 in HEK293 cells

Inducible knockout of CD81 in HEK293 cells. Target cells were first transduced with Tet-On 3G lentivirus and selected with G418. 24 independent Tet-On 3G-positive clones were assayed for induction of the luciferase reporter, and one clone exhibiting the lowest residual expression was selected and transduced with Cas9 lentivirus at an MOI of 5. Target cells were selected with 1 µg/ml puromycin for two weeks, and six stable clones were randomly selected for analysis of Cas9 induction. Cells from each clone were divided between two wells of a 12-well plate and incubated either in the presence or absence of doxycycline (0.5 µg/ml) for two days. Cas9 induction was then assayed by Western blot using polyclonal anti-Cas9 antibody (Cat. No. 632607), diluted 1:1,000, and ECL reagent. Cells derived from three of the Tet-On 3G-Cas9-positive clones (C-1, C-2, and C-3) were then divided between two wells of a 12-well plate and transduced with CD81-sgRNA lentivirus at an MOI of 5. Eight hours after transduction, doxycycline was added to one well for each clone at a concentration of 0.5 µg/ml, and cells were cultured for six days. Cells were then harvested and stained with a FITC-tagged anti-human CD81 antibody and scanned for CD81 expression by FACS. Panel A. Western blot results for six independent Tet-On 3G-Cas9-positive cell populations cultured in the absence (–) or presence (+) of 0.5 µg/ml doxycycline, assayed with anti-Cas9 antibody. Cas9 expression is either undetectable or highly elevated in the absence or presence of doxycycline, respectively. Panel B. FACS results for three independent Tet-On 3G-Cas9-positive 293T cell populations transduced with CD81-sgRNA lentivirus and cultured in the absence (–Dox) or presence (+Dox) of 0.5 µg/ml doxycycline. Two of the populations (C-1 and C-2) exhibited no residual editing in the absence of doxycycline, and knockout of CD81 in either 58.6% or 30.7% of cells, respectively, while the third population (C-3) exhibited residual editing (27.9% of cells) in the absence of doxycycline.

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Inducible knockout of CD81 in Jurkat cells

Inducible knockout of CD81 in Jurkat cells
Inducible knockout of CD81 in Jurkat cells. Tet-On 3G-Cas9-positive Jurkat cells with low residual Cas9 expression were generated using the established protocol and transduced twice with CD81-sgRNA lentivirus at an MOI of 5 on successive days. Cells were then divided between two wells of a 12-well plate and cultured in the absence (–Dox) or presence (+Dox) of doxycycline at a concentration of 0.5 µg/ml for seven days. Cells were then treated with FITC anti-human CD81 antibody and analyzed by FACS. Only a small proportion of cells (0.4%) cultured in the absence of doxycycline (–Dox; top) exhibit residual editing. A significant proportion of cells (32.0%) cultured in the presence of doxycycline (+Dox; bottom) exhibit knockout of CD81.

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Inducible editing of AAVS1 in HEK293 cells

Inducible editing of AAVS1 in HEK293 cells

Inducible editing of AAVS1 in HEK293 cells. Induction of Cas9 expression in six stable Tet-On 3G Cas9-positive HEK293 cell lines was assayed by qRT-PCR. Cells from each line were then divided between two wells of a 12-well plate and transduced with AAVS1-sgRNA lentivirus at an MOI of 5. Eight hours after infection, doxycycline was added to one well for each cell line at a concentration of 0.5 µg/ml, and the cells were cultured for six days. The Guide-it Mutation Detection Kit (Cat. # 631443) was then used to screen for editing of AAVS1. Panel A. qRT-PCR results for Cas9 expression in uninduced (–Dox) and induced (+Dox) cells for each clone. Ct and ΔCt values for uninduced and induced clones are indicated in the corresponding columns, and the calculated fold-difference in expression for each clone is indicated in the column on the right. Of the various clones tested, Clone 1 (numbers in green) exhibited the highest-fold inducibility, while only Clone 3 (numbers in red) exhibited residual expression of Cas9 in the absence of doxycycline and low inducibility. Panel B. Results of Guide-it Resolvase assay for detection of AAVS1 gene editing in uninduced (–) and induced (+) clones. In contrast with the assay of the parental line, which yielded bands of equivalent size for uninduced and induced cells, assays of the induced clones yielded additional bands of smaller sizes, indicating the occurrence of gene editing at AAVS1 for all clones tested. Cleavage products are also visible for the uninduced cells from Clone 3, consistent with the residual expression of Cas9 observed in the qRT-PCR assay.

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Vector components for Lenti-X Tet-On 3G CRISPR/Cas9 System

Vector components for Lenti-X Tet-On 3G CRISPR/Cas9 System

Vector components for Lenti-X Tet-On 3G CRISPR/Cas9 System. pLVX-EF1a-Tet3G encodes the Tet-On 3G transactivator protein under the control of the constitutive EF-1 alpha promoter. In contrast with the commonly used CMV promoter, EF-1 alpha is less susceptible to silencing by certain cell types (including various types of stem cells), enabling the establishment of inducible cell lines that are suitable for long-term use. Tet-On 3G is a modified form of the Tet-On Advanced transactivator protein, which was evolved to have a far higher sensitivity to doxycycline. pLVX-TRE3G-Cas9-puro encodes a modified version of the Cas9 endonuclease under the control of the inducible PTRE3GV promoter. The Cas9 sequence in pLVX-TRE3G-Cas9-puro was originally derived from the bacterium S. pyogenes and has been codon optimized for expression in mammalian cells. Additional Cas9 modifications include the incorporation of a C-terminal nuclear localization signal (NLS). pLVX-hyg-sgRNA1 encodes an sgRNA (designed and inserted by the user) under the constitutive human U6 promoter.

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Choosing an optimal clone for inducible genome editing

Choosing an optimal clone for inducible genome editing

Choosing an optimal clone for inducible genome editing. Clones may be selected by assaying Cas9 expression or genome editing in both uninduced and induced cell populations. However, while Western blot detection of Cas9 is helpful for prescreening clones, CRISPR/Cas9-mediated editing is so efficient that even when Cas9 protein is undetectable by Western blot, genome editing can still occur in corresponding clones. We recommend qRT-PCR for identifying clones in which the residual expression of Cas9 is low enough for editing to be avoided in the absence of doxycycline. Data in the upper row is from a desirable clone (C-1), which exhibits robust expression of Cas9 protein in induced cells (Western blot, +), and the lowest residual levels of Cas9 mRNA (qRT-PCR). Genome editing occurs at much higher frequencies upon induction of Cas9 expression in cells derived from this clone (+ or +Dox) relative to uninduced cells (– or –Dox), as evidenced by the increase in smaller-sized bands in the resolvase assay (blue arrows) and the greater proportion of cells in the “knockout” category as determined by FACS, with the lowest residual editing in uninduced cells. In contrast, data in the lower row is from an undesirable clone (C-3), which exhibits residual transcription of Cas9 (qRT-PCR) and detectable frequencies of genome editing (Resolvase, FACS) in the absence of doxycycline, even though Cas9 protein expression in uninduced cells (–) is undetectable by Western blot

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632633: Lenti-X Tet-On 3G CRISPR/Cas9 System

632633: Lenti-X Tet-On 3G CRISPR/Cas9 System
632629 Lenti-X™ CRISPR/Cas9 System 1 System USD $1144.00

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The Lenti-X CRISPR/Cas9 System is a complete systems for lentiviral-mediated CRISPR/Cas9 genome editing. Cells transduced with this system will constitutively express Cas9 under the CMV promoter and sgRNA under the U6 promoter. We found that transducing the Cas9 and sgRNA separately resulted in much higher editing efficiency, so this system provides the Cas9 and sgRNA on separate vectors. The sgRNA vector is pre-linearized for easy insertion of your guide sequence and the system contains enough pre-linearized vector for 10 separate reactions.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Either Jurkat or HT1080 cells were first transduced with LVX-hyg-CD81-sgRNA and then selected for stable integration using hygromycin

Either Jurkat or HT1080 cells were first transduced with LVX-hyg-CD81-sgRNA and then selected for stable integration using hygromycin
Either Jurkat or HT1080 cells were first transduced with LVX-hyg-CD81-sgRNA and then selected for stable integration using hygromycin. Stable clones were then transduced with LVX-puro-Cas9. Stable clones were selected for using puromycin and then screened for CD81 knockout efficiency using FACS. Positive and negative controls of parental cells without Cas9 transduction were done either with or without anti-CD81 antibody.

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632629: Lenti-X CRISPR/Cas9 System

632629: Lenti-X CRISPR/Cas9 System

*You must be logged in to a Purchasing Account in order to purchase these products online, since the purchase of these products may be restricted depending on your account type. Researchers at not-for-profit accounts receive a limited use license with their purchase of the product. Researchers at for-profit accounts must obtain a license prior to purchase. For details please contact licensing@takarabio.com.

Overviews Lenti-X Concentrator overview
Videos About Lenti-X packaging
Tips and FAQs Lenti-X GoStix Plus FAQs

Overview

  • Viral delivery of the Cas9 gene and a sgRNA allows for genome editing in hard-to-transfect mammalian cells, including proliferating and non-proliferating cells
  • Lenti-X Packaging Single Shots make generating high-titer lentivirus extremely simple
  • Separate systems available for either constitutive or inducible Cas9 expression in target cells
  • Tightest possible control of Cas9 expression (Lenti-X Tet-On 3G CRISPR/Cas9 System) allows you to minimize the likelihood of toxicity and off-target effects associated with persistent Cas9 expression in cell culture
  • Learn about phenotypic screening using a lentiviral whole genome CRISPR library.

More Information

Applications

  • Lentivirus-based delivery of a user-defined sgRNA and Cas9 for mammalian genome editing using CRISPR/Cas9 technology

Components

  • Lenti-X CRISPR/Cas9 System (Cat. # 632629)

    • pLVX-hyg-sgRNA1 Vector (Linear)
    • pLVX-puro-Cas9 Vector
    • Stellar Competent Cells
    • Guide-it Ligation Components v2
    • Lenti-X Packaging Single Shots
  • Lenti-X Tet-On 3G CRISPR/Cas9 System (Cat. # 632633)

    • pLVX-hyg-sgRNA1 Vector (Linear)

    • pLVX-TRE3G-Cas9-puro Vector Set

    • pLVX-EF1a-Tet3G Vector

    • Stellar Competent Cells

    • Guide-it Ligation Components v2

    • Lenti-X Packaging Single Shots

  • pLVX-hyg-sgRNA1 Vector System (Cat. # 632630)

    • pLVX-hyg-sgRNA1 Vector (Linear)

    • Guide-it Ligation Components v2

    • Stellar Competent Cells

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Components List to determine kit components. Certificates of Analysis and Components Lists are located under the Documents tab.


Lenti-X Concentrator

Lenti-X Concentrator video

Lenti-X Concentrator is a simple and fast method for concentrating any lentiviral stock—just mix your lentiviral supernatant with the concentration reagent, incubate, and spin the mixture in a standard centrifuge. Watch the video to see how it works.

Watch Concentrator video Lentiviral packaging overview

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