TARGATT™ Site-Specific Knock-in Cell Line Service

TARGATT™ Fast and Site-Specific Gene Insertion in Mammalian Cell Lines
 
Do you need a precise comparison of different genes from more than one cell line? Do you want to generate a master cell line expressing different reporter genes? TARGATT™ provides transgene expression and a knock-in cell line in one step.
 
Generation of TARGATT master knock-in cell line with attP docking sites allows:
  • Site-specific integration at pre-defined loci.
  • High integration efficiency with no disruption of endogenous genes
  • Single-copy integration and stable expression
  • Single-step transfection to create stable cell lines 
  • No massive clone screening required
  • Inclusion of large transgenes up to 20 kb
Applied StemCell is now providing a TARGATT master knock-in cell line generation service
  • You send us your favorite cell lines
  • We will insert the attP docking sites at specific loci
  • Fast: get your transgenic cell line in 3 months!
Need Site-Specific Knock-in of iPSCs? Applied StemCell can do that too!
 
Site-Specific Knock-in of TARGATT™ sites
Knock In Cell Line Cell Line Generation Service
Figure: Using mCherry reporter (left) in human iPSC line. Original Cell Line: hiPSC (right).

FAQ

FAQ for Gene Knock-in Technology and TARGATT™ Master Knock-in Cell Line Service

1. I am interested in making a master TARGATT™ cell line using my own cell line as a parental cell line. Can you take my cancer cell line, for example, to generate a master cell line using your service?

Yes.

2. What is the Gene Knock-in Technology used to generate master cell lines?

We use CRISPR/Cas9 to generate master cell lines by inserting TARGATT™ attP sites at a desired locus.

3. How long does it take to make a master cell line?

3-5 months, depending on the nature of the cell line.

4.  Is there a size limit on DNA to be inserted into the genome (to attP site)?

So far we have successfully inserted a transgene of 20kb using this method.

5. What is the final deliverable product?

We ship at least 2 vials, each at 0.5x10^6 cells/vial, cryopreserved cells per clone with a report of the project. Additional clone(s) and vial(s) are available upon request.  For fee-for service projects, you can also have the CRISPR and TARGATT™ vectors upon request.

6. What safe harbor loci are available to place the TARGATT™ docking site? Can you recommend one for my cell lines?

We have several ready-to-use safe harbor loci to choose from. They are Rosa26, H11, and AAVS1. Docking site insertion can be customized to other loci base on your project.

7. I got my master cell line. Do you provide plasmid so that we can construct Knock-in vector with our gene of interest?

Yes.

8. Do you have off-the-shelf TARGATT™ Master cell lines?

Currently, we have human iPSC and mouse C57/BL6 ESC and iPSC lines. CHO cells will be launched soon. We are happy to discuss for other cell lines as Custom Cell Line Services.

 

References

Description of the technology

Commentary, comparison with other transgenic methods

Tet inducible mice generated by TARGATT™

Advantage of Hipp11 (H11) locus

Application for mice generated by TARGATT™

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