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

Proc Natl Acad Sci U S A. 2017 Jan 24;114(4):722-727. doi: 10.1073/pnas.1615735114. Epub 2017 Jan 6.

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.

Keywords: CRISPR/Cas9; liver cancer; piggyBac transposon; screening; tumorigenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics*
  • Carcinogenesis / genetics*
  • DNA Transposable Elements / genetics
  • Gene Library
  • Genes, Tumor Suppressor / physiology
  • Genetic Engineering / methods
  • Genome / genetics
  • Liver / pathology
  • Liver Neoplasms / genetics
  • Liver Neoplasms / pathology
  • Mice
  • RNA, Guide / genetics

Substances

  • DNA Transposable Elements
  • RNA, Guide