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piggyBac mediates efficient in vivo CRISPR library screening for tumorigenesis in mice
- Source :
- Proceedings of the National Academy of Sciences. 114:722-727
- Publication Year :
- 2017
- Publisher :
- Proceedings of the National Academy of Sciences, 2017.
-
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.
- Subjects :
- 0301 basic medicine
Transposable element
Carcinogenesis
Computational biology
Biology
medicine.disease_cause
Genome
Mice
03 medical and health sciences
In vivo
medicine
Animals
CRISPR
Genes, Tumor Suppressor
Guide RNA
Gene
Gene Library
Genetics
Multidisciplinary
Cas9
Liver Neoplasms
Biological Sciences
030104 developmental biology
Liver
DNA Transposable Elements
CRISPR-Cas Systems
Genetic Engineering
RNA, Guide, Kinetoplastida
Subjects
Details
- ISSN :
- 10916490 and 00278424
- Volume :
- 114
- Database :
- OpenAIRE
- Journal :
- Proceedings of the National Academy of Sciences
- Accession number :
- edsair.doi.dedup.....ffe5485c0a44a4d030b71f2bae6d52c5