In vivo profiling of metastatic double knockouts through CRISPR-Cpf1 screens

Nat Methods. 2019 May;16(5):405-408. doi: 10.1038/s41592-019-0371-5. Epub 2019 Apr 8.

Abstract

Systematic investigation of the genetic interactions that influence metastatic potential has been challenging. Here we developed massively parallel CRISPR-Cpf1/Cas12a crRNA array profiling (MCAP), an approach for combinatorial interrogation of double knockouts in vivo. We designed an MCAP library of 11,934 arrays targeting 325 pairwise combinations of genes implicated in metastasis. By assessing the metastatic potential of the double knockouts in mice, we unveiled a quantitative landscape of genetic interactions that drive metastasis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Bacterial Proteins / genetics*
  • CRISPR-Associated Protein 9 / genetics
  • CRISPR-Cas Systems / genetics*
  • Cell Line, Tumor
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Endonucleases / genetics*
  • Gene Editing / methods*
  • Gene Knockout Techniques / methods*
  • Mice
  • Neoplasm Metastasis / genetics*
  • Sequence Analysis, RNA

Substances

  • Bacterial Proteins
  • CRISPR-Associated Protein 9
  • Cpf1 nuclease, Acidaminococcus
  • Endonucleases