With Cellartis gene editing services, we will deliver high-quality, edited human iPS cells from your disease- or patient-specific donor lines. The cells will be edited according to your specifications using CRISPR/Cas9 technology.
The gene editing services start with you sending us your own human iPS cells. As an alternative, you may request the Cellartis sourcing and reprogramming services. Our gene editing protocol starts with thawing and adapting your pluripotent cells to the Cellartis DEF-CS 500 Culture System, and concurrent design and production of sgRNAs and ssDNA for knockins. The cells are then edited via electroporation-based delivery of ribonucleoprotein (RNP) complexes, screened, and single-cell cloned. Finally, clones of interest are identified, expanded, and characterized. Depending on your downstream applications, you may request the Cellartis directed differentiation services for your edited cells.
Cellartis Human Pluripotent Stem Cell Services
Gene editing
The combination of precise, footprint-free gene editing using CRISPR/Cas9 and human pluripotent stem cells allows for the generation of disease models that are essential for advancing our understanding of disease mechanisms and the development of novel therapeutics. However, creating these models can be extremely challenging as successful gene editing and concurrent establishment of clonal populations can be both inefficient and time-consuming. To help you create your unique disease models, we offer gene editing services to knock out a target gene or knock in a disease-specific mutation. With the help of our services, you can focus on your research and leave the challenges to us.
For more information about our services, please submit the service inquiry form below.
Overview
Thawing of your human iPS cells and adaptation to the DEF-CS system
Project types
Service | What kind | Alleles Affected | Zygosity | Notes | Case study |
Knockout | Out-of-frame knockout (first exon) | monoallelic | heterozygous | Knocking out an endogenous gene (CD81) in hiPS cells | |
biallelic | heterozygous | ||||
Protein truncation | monoallelic | heterozygous | |||
biallelic | heterozygous | ||||
Knockin | SNP | monoallelic | heterozygous |
|
Introducing a tyrosinemia-related SNP in hiPS cells |
biallelic | homozygous | ||||
Gene-tagging (longer sequences up to 4 kb; fluorescent proteins, DYKDDDDK, Myc, etc.) | monoallelic | heterozygous |
|
||
biallelic | homozygous | ||||
Complete expression cassette | monoallelic | heterozygous |
|
Inserting an expression cassette into the AAVS1 locus in hiPS cells | |
biallelic | homozygous |
Deliverables/lead time
Service | Cat. # | Deliverables | Lead time |
Cellartis Human iPS Gene Editing Service (Knockout)* | Y60120 |
|
Approx. 26 weeks |
Cellartis Human iPS Gene Editing Service (Knockin)* |
Y60121 |
|
Approx. 28 weeks |
*If you are interested in gene editing services for human embryonic stem cells, please contact us by submitting the form below.
QC testing
Included with gene editing services:
- Visual inspection and confirmation of cell morphology and proliferation
- Immunocytochemistry to verify the presence of the pluripotency markers Oct-4, SSEA-4, TRA-1-60, and TRA-1-81
- Sequence analysis of both alleles
- Sterility testing of the final cell product (bacteria, including Mycoplasma)
Additional testing and characterization services for edited cells:
- Cellartis Descriptive Karyotyping Service (Cat. # Y60304)
- Karyotype analysis of 20 G-banded metaphase spreads from edited cells will be performed.
- Cellartis Spontaneous In Vitro Differentiation Service (Cat. # Y60305)
- A 3-week spontaneous in vitro differentiation protocol will be used on a subset of edited cells. Cells will be fixed and immunolabelled for ASMA (mesoderm), FoxA2 (endoderm), and βIII-tubulin (ectoderm) to confirm trilineage potential.
- Cellartis STR Analysis Service (Cat. # Y60306)
- Genomic DNA will be isolated from edited cells and the corresponding starting material. Short tandem repeat (STR) analysis will be performed on 16 loci to verify there are no differences between the samples.
Project initiation
You may wish to submit your own human iPS cells for Cellartis gene editing services, or use Cellartis sourcing and reprogramming services to generate the starting material. If you would like to start with your cells, we will need 5 vials (>1 million viable cells per vial) of hiPS cells, with relevant information, including name of the donor cell line, culture conditions, and cell density upon freezing. Your cells will be first adapted to the DEF-CS system, and then the editing protocol can begin.
For safety and QC purposes, we can only work with human samples that are free of human pathogens and that have passed a sterility test. We require documentation proving that all samples shipped to us are sterile and pathogen-free. In the absence of documentation, testing for pathogens (Cat. # Y60301) and sterility (Cat. # Y60302) will be performed via a third-party service prior to the initiation of Cellartis services. Please note that safety and sterility testing will incur additional lead time and costs.
- Submit the service inquiry form below.
- We contact you to go over your project requirements and provide a quote.
- Place your order and ship us the necessary cells and documentation to initiate the service. (Does not apply if using our sourcing service.)
- We perform the agreed-upon service according to the designated lead times.
- We provide you with the agreed-upon deliverables.
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