Overview
- Species: Human
- Features: cryopreserved, female, TARGATT
- Product Type: TARGATT Kits
- iPSC: iPSC control line, iPSC Services
- Features:contact us for more information
TARGATT™ Human Induced Pluripotent Stem Cell (hiPSC) Master Cell Line (Female) Knock-in Kit
Cat.-Nr.: AST-1602
Description
Quickly create the most relevant cell line for your studies with iPSCs that are ready for TARGATT™ knock-in and then differentiation into your desired cell type.
- TARGATTT™ advantages—site-specific, single copy knock-in of 20+ kb of DNA at efficiencies approaching or exceeding 90% after selection
- Gene editing-ready iPSCs—save time with hiPSCs that come with a TARGATT™ landing pad engineered into the H11 safe harbor locus
- iPSC flexibility—differentiate into your desired cell type after integrating your transgene
- Male and female iPSCs available—both generated using episomal factors from well-characterized, karyotype normal iPSCs—find the male kit here
The TARGATT™ hiPSC Master Cell Line (Female) Knock-in Kit from Applied StemCell accelerates your cell line development by providing iPSCs with a pre-engineered TARGATT™ landing pad at the H11 safe harbor locus. Simply clone your DNA of interest into the TARGATT™ cloning plasmid, transfect into the TARGATT™ iPSC Master Cell Line (Female), select for integrants, and differentiate into your desired cell type.
Each AST-1602 contains sufficient cells and plasmids for 3 transfections and comes with:
- TARGATT™ hiPSC Master Cell Line (Female, AST-1602-C)—engineered with a TARGATT™ landing pad at the H11 safe harbor locus of ASE-9211 hiPSCs (episomally reprogrammed fibroblasts, 47-year-old Caucasian female donor).
- TARGATT™ 45 CAG-MCS Cloning Plasmid (AST-3086)
- TARGATT™ 44 mCherry-Blas Control Plasmid (AST-3085)
- TARGATT™ CMV-integrase Plasmid (AST-3084)
With the large-insert capabilities of the TARGATT™ platform, you can easily and efficiently insert large genes, multiple genes, and complex constructs—all types of editing that CRISPR/Cas9 is unable to achieve in a single reaction.
Integration is specific for sequences in the TARGATT™ landing pad, which we’ve pre-engineered into a well-characterized intergenic region—the H11 safe harbor locus—in our hiPSC Master Cell Lines. Gene expression from the H11 locus is consistent and robust, resulting in reliable protein production. In addition, because TARGATT™-mediated integration is unidirectional and irreversible, unlike insertion with cre-lox and flp-frt systems, gene expression is stable over time and with each passage.
SUPPLIER:
Applied StemCell
STATUS:
In Stock
SIZE:
kit
FAQs
Can I use the TARGATT™ iPSC Master Cell Line for clinical applications?
No, the TARGATT™ iPSC Master Cell Line is for research use only. We offer GMP-grade TARGATT™ iPSCs (AST-9480) for clinical applications.
How was the TARGATT™ iPSC Master Cell Line created?
The TARGATT™ iPSC Master Cell Line is engineered from ASE-9211, a well-characterized, karyotype-normal iPSC line. ASE-9211 was reprogrammed from fibroblasts using episomal reprogramming.
How does TARGATT™ technology compare to other gene insertion methods?
TARGATT™ technology enables highly efficient, single-copy, site-specific integration at the H11 safe harbor locus. Integration is stable and results in long-term expression—advantages that other random or multi-copy insertion methods cannot consistently provide.
What are the best applications for the TARGATT™ iPSC Master Cell Line?
TARGATT™ technology is ideal for:
- Reporter knock-in cell lines
- Drug discovery screening
- Library screening
- Large transgene knock-ins for therapeutic studies
What is the knock-in efficiency of TARGATT™ technology?
TARGATT™ technology provides high knock-in efficiency with stable and site-specific gene integration. After drug selection, efficiency can approach 90%.
What reagents or materials are required for the TARGATT™ process but not included in the kit?
The following materials are not included and must be procured separately:
| Material | Vendor | Catalog Number |
|---|---|---|
|
mTeSR™ Plus
|
Stem Cell Technologies
|
05825
|
|
Matrigel® hESC-Qualified Matrix
|
Corning
|
354277
|
|
Rock Inhibitor Y-27632
|
Segment
|
04-0012-02
|
|
CryoStor® CS10 Freeze Media
|
Stem Cell Technologies
|
210102
|
|
0.5mM EDTA in PBS
|
Life Technologies
|
15575-020
|
|
TrypLE™ Express Enzyme (1X)
|
Gibco
|
12604013
|
|
Neon™ Transfection System Kit
|
ThermoFisher
|
MPK10096
|
|
Hygromycin
|
InvivoGen
|
Antihg-1
|
|
Penicillin-Streptomycin (100X)
|
Gibco
|
15140-122
|
|
Ganciclovir (GCV)
|
Selleck Chemicals
|
S1878
|
If you are using similar or alternative reagents, we recommend ensuring they align with the TARGATT™ culture protocols for optimal results.
How does TARGATT™ ensure minimal off-target effects?
The TARGATT™ system uses a patented serine integrase that specifically recognizes TARGATT attB sites in the cloning plasmid (also referred to as the donor plasmid) and attP sites in the TARGATT landing pad. Because unbound double strand breaks are not created, random integration using the host double strand break repair machinery does not occur.
What is the cargo size capacity of TARGATT™ technology?
TARGATT™ can efficiently insert payloads of 20 kb in a single reaction and larger payloads with nested TARGATT™ landing pads, making the technology suitable for a wide range of genetic engineering applications.
Can I customize the gene of interest for knock-in using TARGATT™?
Yes, the TARGATT™ cloning plasmid allows you to clone and amplify your desired gene of interest for site-specific insertion.
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