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Guide-it Recombinant Cas9 (Electroporation-Ready)
Overviews Adoptive T-cell therapy with RetroNectin GMP grade reagent
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Guide-it Recombinant Cas9 (Electroporation-Ready)
Overviews Adoptive T-cell therapy with RetroNectin GMP grade reagent

GMP recombinant Cas9

GMP recombinant Cas9

Recombinant Cas9 Protein GMP grade is a wild-type S. pyogenes Cas9 nuclease fused with a C-terminal NLS for use in gene editing experiments. It was designed without his-tag sequences, making it appropriate for use in various applications.

This reagent is manufactured as a quality-assured product according to relevant guidelines for Good Manufacturing Practice (GMP) and is suitable for ex vivo cell engineering and gene therapy studies. Please complete and return this form prior to purchase.

Cat. # Product Size Price License Quantity Details
T230 Recombinant Cas9 Protein GMP grade 0.6 mg USD $3211.00

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.

Recombinant Cas9 Protein GMP grade, is a recombinant wild-type Streptococcus pyogenes Cas9 nuclease with a nuclear localization signal (NLS) located at the C-terminus of the protein. It is manufactured and tested in accordance with the following guidelines: "Standards for Manufacturing Control and Quality Control, etc. of Investigational Products (Investigational products GMP)," as notified by the Ministry of Health and Welfare Japan ("PFSB Notification No. 0709002, July 9, 2008"; PFSB = Pharmaceutical and Food Safety Bureau). No animal- or human-derived components are used in the manufacture of this product. The Cas9 protein solution has been verified to be sterile and well-tolerated by mammalian cells when electroporated as a ribonucleoprotein complex (RNP) with a single-guide RNA (sgRNA) for knockout experiments, or as an RNP with a donor repair template for knockin experiments.

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.

Documents Components Image Data

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Knockout of cell-surface proteins in human iPS cells.

Knockout of cell-surface proteins in human iPS cells.

Knockout of cell-surface proteins in human iPS cells. In this experiment, human iPS cells were electroporated with either competitors' Cas9 (Vendor X and Vendor Y)/sgRNA RNPs or Recombinant Cas9 Protein GMP grade (Cat. # T230)/sgRNA RNPs against cell-surface protein A (gene A) and/or protein B (gene B). Cells were stained with either anti-protein A antibody or anti-protein B antibody and then run on a FACS machine to detect each surface protein. Takara Bio's GMP-grade Cas9 protein showed high knockout efficiency at each condition tested. Data provided by Drs. Akitsu Hotta and Huaigeng Xu at the Center for iPS Cell Research and Application (CiRA), Kyoto University.

Back

Knockout of CD81 in 293T/17 cells.

Knockout of CD81 in 293T/17 cells.

Knockout of CD81 in 293T/17 cells. In this experiment, 293T/17 cells were electroporated with either Guide-it Recombinant Cas9 (Electroporation-Ready; Cat. # 632641)/sgRNA RNPs or Recombinant Cas9 Protein GMP grade (Cat. # T230)/sgRNA RNPs against CD81. Electroporation was performed at 1,300, 1,200, and 1,100 V for conditions #1, 2, and 3, respectively. Pulse interval was 20 ms, for each of two pulses, for each condition. Cells were stained with CD81 PE-labeled anti-CD81 antibody (BioLegend, Inc.; Cat. # 349506) and then run on a FACS machine to detect CD81 seven days after editing. The negative staining control shows the FACS data from cells without antibody staining, while the electroporation control shows cells electroporated without Cas9/sgRNA RNPs. When cells were electroporated with RNPs for both Guide-it Cas9 (632641) and GMP Cas9 (T230), high editing efficiencies were observed.

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T230: Recombinant Cas9 Protein GMP grade

T230: Recombinant Cas9 Protein GMP grade

Overview

Recombinant Cas9 Protein GMP grade, is manufactured and tested in accordance with the following guidelines: "Standards for Manufacturing Control and Quality Control, etc. of Investigational Products (Investigational products GMP)," as notified by the Ministry of Health and Welfare Japan ("PFSB Notification No. 0709002, July 9, 2008)"; PFSB = Pharmaceutical and Food Safety Bureau). 

  • No animal- or human-derived components are used in the manufacture of this product
  • No his-tag sequence is in this product—appropriate for various applications; researchers recently reported a first in-human phase 1 clinical trial using engineered T cells edited by a different untagged recombinant Cas9 (Stadtmauer et al 2020)
  • Appropriate for gene editing in various cell types and organisms when electroporated or microinjected as a ribonucleoprotein complex (RNP) with a single-guide RNA (sgRNA); please complete and return this form prior to purchase

More Information

Applications

Gene editing

References

Jinek, M. et al. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. 337, 816 (2012).

Liang, X. et al. Rapid and highly efficient mammalian cell engineering via Cas9 protein transfection. J. Biotechnol. 208, 44 (2015).

Sander, J.D. & Joung, J.K. CRISPR-Cas9 systems for genomic editing, regulation and targeting. Nat. Biotechnol. 32, 347 (2014).

Stadtmauer, E.A. et al. CRISPR-engineered T cells in patients with refractory cancer. Science. 367, eaba7365 (2020).

Additional product information

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


Tech note describing the use of RetroNectin and CD3 antibody for

T-cell expansion

RetroNectin Recombinant Human Fibronectin Fragment and RetroNectin GMP grade have widespread use as enhancers of retroviral or lentiviral gene transfer into hard-to-transduce cell types. Here, we present data on an additional application—the improved expansion of T cells from PBMCs and increased production of naïve T cells—using a combination of our products: GT-T551 T-cell medium, RetroNectin reagent, CultiLife culture bags, and anti-CD3 antibody.

T-cell expansion tech note Related products

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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