Any success in generating KI mice through microinjecting TALEN mRNA + dsDNA ?

[Tsai Stevie]

Dear List,

We've been trying to introduce EGFP into a mouse gene loci through TALEN by co-injecting TALEN pair RNA and a dsDNA(~2.1kb EGFP cDNA floxed with 1 and 4 kb homologous arms) as donor.

With the locus-specific TALEN alone, we can get around 70% Indel from the pups, however, plus the donor DNA, we cannot get any HRD-offsprings.

Is there anyone using dsDNA as HR donor instead of ssODN?

Any success?

Thank you in advance.

Ching-Yen Ysai, Ph.D.

Manager, Transgenic Core Facility

Academia Sinica, TAIWAN