Gene knock-in

Knock-in of a Large Reporter Gene via the High-Throughput Microinjection of the CRISPR/Cas9 System
Knock-in of a Large Reporter Gene via the High-Throughput Microinjection of the CRISPR/Cas9 System 780 435 IEEE Transactions on Biomedical Engineering (TBME)
This work demonstrates the microinjection of CRISPR/Cas9 with an enhanced green fluorescent protein (GFP) donor template into single HepG2 cells can achieve reporter gene knock-in targeting the adeno-associated virus site 1 locus. Homology-directed repair-mediated knock-in can be observed with an efficiency of 41%. Assessment via T7E1 assay indicates that the GFP knock-in cells exhibit no detectable changes at potential off-target sites. A case study of injecting the GFP knock-in cells into zebrafish (Danio rerio) embryos to form an in vivo tumor model is conducted. Results demonstrate the efficiency of combining microinjection with the CRISPR/Cas9 system in achieving gene knock-in. read more
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