A Novel Approach for Making Knockin Zebrafish Without Disrupting Endogenous Targeted Genes

I am Jiu-lin Du at the Institute of Neuroscience, Chinese Academy of Sciences. Recently, my lab develops an intron targeting-mediated knockin approach, by which we can efficiently generate knockin zebrafish lines. The paper is just online at Cell Research (http://www.nature.com/cr/journal/vaop/ncurrent/pdf/cr201543a.pdf).

Using this method, we can insert a large exogenous DNA sequence (GFP, Gal4 et al.) into the zebrafish genome without interrupting the coding sequence and the Cis-regulatory elements of the targeted gene. Therefore, the exogenous gene can well recapitulate the expression pattern of endogenous targeted genes, and the expression and function of the targeted gene will not be affected.

I think it may be helpful for our society. If you are interested in the details, please don't hesitate to contact us. forestdu@ion.ac.cn

 Li, J., Zhang, B.B., Ren, Y.G., Gu, S.Y., Xiang, Y.H., Huang, C., and Du, J.L. (2015). Intron targeting-mediated and endogenous gene integrity-maintaining knockin in zebrafish using the CRISPR/Cas9 system. Cell research (Online).