title: piggyBac mediates efficient in vivo CRISPR library screening for tumorigenesis in mice.
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 in delivery. Here, we examined 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.
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Chunlong
authors.initials: C
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Xiaolan
authors.initials: X
authors.lastname: Qi
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Xuguang
authors.initials: X
authors.lastname: Du
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Huiying
authors.initials: H
authors.lastname: Zou
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Feng
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Hengxing
authors.initials: H
authors.lastname: Lu
authors.affiliation: Department of Neurosurgery, Medical School, University of Texas Health Science Center at Houston, Houston, TX 77030.|Center for Stem Cell and Regenerative Medicine, University of Texas Health Science Center at Houston, Houston, TX 77030.
authors.forename: Shenglan
authors.initials: S
authors.lastname: Li
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: An
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Human Genetics, University of Utah School of Medicine, Salt Lake City, UT 84112.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Neurosurgery, Medical School, University of Texas Health Science Center at Houston, Houston, TX 77030.|Center for Stem Cell and Regenerative Medicine, University of Texas Health Science Center at Houston, Houston, TX 77030.|The Senator Lloyd & B. A. Bentsen Center for Stroke Research, the Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases, University of Texas Health Science Center at Houston, Houston, TX 77030.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Li
authors.affiliation: Department of Human Genetics, University of Utah School of Medicine, Salt Lake City, UT 84112; swu@cau.edu.cn capecchi@genetics.utah.edu.
authors.forename: Mario R
authors.initials: MR
authors.lastname: Capecchi
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China; swu@cau.edu.cn capecchi@genetics.utah.edu.
authors.forename: Sen
authors.initials: S
authors.lastname: Wu
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D063646:Carcinogenesis / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics; D015723:Gene Library; D016147:Genes, Tumor Suppressor / Q000502:physiology; D005818:Genetic Engineering / Q000379:methods; D016678:Genome / Q000235:genetics; D008099:Liver / Q000473:pathology; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology; D051379:Mice; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics
chemical_list: D004251:DNA Transposable Elements; D000094704:RNA, Guide, CRISPR-Cas Systems
keywords: CRISPR/Cas9; liver cancer; piggyBac transposon; screening; tumorigenesis