Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems  

作　　者：Yuqian Jiang Rachel Catherine Hoenisch Yun Chang Xiaoping Bao Craig E.Cameron Xiaojun Lance Lian 

机构地区：[1]Department of Biomedical Engineering,Pennsylvania State University,University Park,PA,16802,USA [2]Huck Institutes of the Life Sciences,Pennsylvania State University,University Park,PA,16802,USA [3]Department of Biology,Pennsylvania State University,University Park,PA,16802,USA [4]Davidson School of Chemical Engineering,Purdue University,West Lafayette,IN,47907,USA [5]Department of Microbiology and Immunology,University of North Carolina School of Medicine,Chapel Hill,NC,27599,USA

出　　处：《Bioactive Materials》2022年第8期313-320,共8页生物活性材料（英文）

基　　金：supported by NIH R21EB026035(X.L.L.);NIH R21AI149312(C.E.C.,and X.L.L.);NSF CBET-1943696(X.L.L.);Penn State startup funding to X.L.L.

摘　　要：CRISPR/Cas-mediated genome editing in human pluripotent stem cells(hPSCs)offers unprecedented opportunities for developing in vitro disease modeling,drug screening and cell-based therapies.To efficiently deliver the CRISPR components,here we developed two all-in-one vectors containing Cas9/gRNA and inducible Cas13d/gRNA cassettes for robust genome editing and RNA interference respectively.These vectors utilized the PiggyBac transposon system,which allows stable expression of CRISPR components in hPSCs.The Cas9 vector PB-CRISPR exhibited high efficiency(up to 99%)of inducing gene knockout in both protein-coding genes and long non-coding RNAs.The other inducible Cas13d vector achieved extremely high efficiency in RNA knockdown(98%knockdown for CD90)with optimized gRNA designs.Taken together,our PiggyBac CRISPR vectors can serve as powerful toolkits for studying gene functions in hPSCs.

关 键 词：CRISPR-Cas9 Genome editing Cas13d RNA editing PiggyBac transposon Human pluripotent stem cells 

分 类 号：Q78[生物学—分子生物学] R318[医药卫生—生物医学工程]