single nucleotide base change

[lacr...@gmail.com]

Hi,

I am new to the Talen technology and I am interested in using this technology to introduce single nucleotide polymorphism in the transcription site of my gene of interests in primary cells. I was wondering if the repair template used for homologous recombination repair will be targeted by the talens? If yes, then how does one avoid the problem? Secondly, how efficient is it to target primary cells? Thanks for your feedback

Warmest regards
Tuang

[Le Cong]

Hi Tuang,

If you could make some silent or non-deleterious mutations to your repair oligo, then you could use this strategy to minimize template cleavage by TALENs. Alternatively, you could design your template to not include both target sites of your TALEN pair, e.g. only have a template with left TALEN site, thereby reducing the possibility of cleaving your template. 

Best,

Le

Le CONG, Ph.D. 丛乐

Broad Institute of MIT and Harvard,

Massachusetts Institute of Technology,

415 Main Street, Cambridge, MA 02142

Tel: (+1) 617-714-7879

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E-Mail: congl...@gmail.com / co...@broadinstitute.org / con...@mit.edu

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Best,

Le Cong