Chunlong Xu,Xiaolan Qi,Xuguang Du,Huiying Zou, Mario R. Capecchi,and Sen Wu.piggyBac mediates efficient in vivo CRISPR library screening for tumorigenesis in mice.PNAS.DOI:10. 1073/pnas.1615735114
发布日期:2019-11-08 浏览次数:  信息来源:生物学院

piggyBac mediates efficient in vivo CRISPR library screening for tumorigenesis in mice


Chunlong Xu, Xiaolan Qi, Xuguang Du, Huiying Zou, Fei Gao, Tao Feng, Hengxing Lu, Shenglan Li, Xiaomeng An, Lijun Zhang, Yuanyuan Wu, Ying Liu, Ning Li, Mario R. Capecchi, and Sen Wu


PNAS.

DOI:10. 1073/pnas.1615735114  44,s170579,吴森.pdf


Abstract

CRISPR/Cas9 is becoming an increasingly important tool to functionally annotate genomes. However, because genome-wide CRISPR libraries are mostly constructed in lentiviral vectors, in vivo applications are severely limited as a result of difficulties indelivery .Here,weexamined the piggyBac (PB)transposon as an alternative vehicle to deliver a guide RNA(gRNA)library for in vivo screening.Although tumor induction has previously been achieved in mice by targeting cancer genes with the CRISPR/Cas9 system, in vivo genome-scale screening has not been reported. With our PB-CRISPR libraries, we conducted an in vivo genome-wide screen in mice and identified genes mediating liver tumorigenesis, including known and unknown tumor suppressor genes (TSGs). Our results  demonstrate that PB can be a simple and nonviral choice for efficient in vivo delivery of CRISPR libraries.


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