Mammalian Cell Line Engineering Services
Creating stable cell lines is time-consuming and complex. The trusted, experienced developers of GeneArt® Precision TALs and GeneArt® CRISPRs can custom-design stable cell lines generated using one of the most robust and reliable technologies on the market. Utilizing quality products throughout the process—everything from Gibco® cell culture media and reagents and cell health assays, to next-generation sequencing using Ion Torrent™ sequencers—our scientists will work with you to design your stable cell line and perform quality control testing to help ensure that the cell line meets your requirements.
Comparison of the CRISPR and TAL effector editing technologies
|What is it?||A DNA-cleaving protein guided by an RNA molecule that is able to modify a specific gene of interest||A protein containing a DNA-cleaving activity and a DNA-binding region that can be programmed to recognize any gene of interest|
|Pros||Pros: This technique is affordable and easy to use, and it works for high-throughput, multigene experiments
||Pros: This technology is simpler and cheaper to customize for specific targets than other technologies, and typically has less off-target cleavage
|Cons||Cons: It can cleave off-target sites||Cons: Constructs are more difficult to produce and deliver|
GeneArt® Precision TALs or GeneArt® CRISPRs are efficient technologies for precise genome modifications in mammalian cells; the table below highlights just a few key gene-editing applications. Speak with our services expert for free consultation in designing the right cell line for your research.
|Stable cell line generation service||Description||Genome editing tool||Donor DNA synthesis|
|Knock-In Stable Cell Line Generation||Add tags (e.g., luciferase, GFP) to your gene of interest; introduce/correct point mutations; safe harbor site (AAVS1)||TAL or CRISPR||Required|
|Knock-Out Stable Cell Line Generation||Knock out endogenous gene or introduce deletions||TAL or CRISPR||Recommended|
|Activator Stable Cell Line Generation||Activate/up-regulate endogenous gene expression||TAL or CRISPR||N/A|
|Repressor Stable Cell Line Generation||Repress/down-regulate endogenous gene expression||TAL or CRISPR||N/A|
What the service includes
We will apply our expertise in creating stable cell lines to design, develop, and validate a custom stable cell line as outlined below, using validated GeneArt® Precision TALs or GeneArt® CRISPR and a customer-supplied cell line.
- Protocol transfer and transfection optimization service
- Custom GeneArt® Precision TALs or GeneArt® CRISPR production
- Validation of custom GeneArt® Precision TALs or GeneArt® CRISPR efficacy
- Stable cell line generation
- Quality control analysis of stable cell line
Process and timelines
How rapidly we can deliver a custom stable cell line depends on the individual cell line growth characteristics and culturing requirements.
Projects are typically completed within 23–30 weeks.
Process for creating GeneArt® Precision TALs or GeneArt® CRISPR stable cell lines.
Q: How do you compare the CRISPR/Cas9 system with TALENs in terms of specificity for the target?
A: It is generally believed that TALENs have higher specific cleavage of the target than CRISPRs. Carefully designing the CRISPR target typically results in lower off-target effects. The cleavage efficiency of CRISPR or TALEN for the target locus can be measured using the GeneArt® Genomic Cleavage Detection Kit.
Q: Why would one use CRISPR as a screening tool before going to TALs?
A: Since cleavage efficiency at a particular locus depends on the accessibility of the locus, chromatin state, and sequence, it is advisable to test multiple different loci/regions within a gene of interest. With CRISPR/Cas9–mediated genome editing, for each target of interest the user needs only to change the 19–20 bp target-specific oligo. After targets have been screened and the sequence/locus with best cleavage efficiency has been identified with the fast and easy-to-use GeneArt® CRISPR system, the biologically relevant mutations can be precisely created with high-specificity GeneArt® Precision TALs.
Q: What are the advantages/disadvantages of OFP vs. CD4 in the GeneArt® CRISPR Nuclease Vectors?
A: One could perform FACS-based sorting with OFP (orange fluorescent protein); either FACS or bead-based enrichment can be used for CD4. Find more details in the user manual.
How to order
To discuss your service project, obtain a Services Quotation, or to order, please contact our Custom Services department:
- Phone (toll-free in North America): 800 955 6288 (extension 4)
- Fax (in North America): 716 774 3157
- Email: firstname.lastname@example.org
- CRISPR Product Flyer
- CRISPR Technical Product Bulletin
- GeneArt® Precision TALs Flyer
- GeneArt Precision TALs User Manual
- Application Note: Improve genome editing outcomes in biologically relevant cell models
- Genome Editing eBook
- Genome Engineering Technology Selection Guide
- Genome Editing Publications
- View our library of Cell Engineering & Genome Editing webinars
Video on TALs technology
Watch this online tutorial, presented by Dr. Jon Chesnut, Research Fellow and R&D lead, to learn more about GeneArt® Precision TALs technology.
Terms and Conditions for Provision of Life Technologies Custom Solutions govern the service work provided.One or more Limited Use Label Licenses may apply to Client's use of deliverables described. The full text of Life Technologies’ Limited Use Label Licenses is found in Life Technologies’ Life Science Products and Services Catalogue or on the web at www.lifetechnologies.com/Support
For Research Use Only. Not for use in diagnostic procedures.