A CRISPR Resource for Individual, Combinatorial, or Multiplexed Gene Knockout

Mol Cell. 2017 Jul 20;67(2):348-354.e4. doi: 10.1016/j.molcel.2017.06.030.

Abstract

We have combined a machine-learning approach with other strategies to optimize knockout efficiency with the CRISPR/Cas9 system. In addition, we have developed a multiplexed sgRNA expression strategy that promotes the functional ablation of single genes and allows for combinatorial targeting. These strategies have been combined to design and construct a genome-wide, sequence-verified, arrayed CRISPR library. This resource allows single-target or combinatorial genetic screens to be carried out at scale in a multiplexed or arrayed format. By conducting parallel loss-of-function screens, we compare our approach to existing sgRNA design and expression strategies.

Keywords: CRISPR; Cas9; algorithm; combinatorial; expression; lentivirus; library; prediction; screen; sgRNA.

Publication types

  • Comparative Study

MeSH terms

  • Algorithms
  • CRISPR-Associated Proteins / genetics*
  • CRISPR-Associated Proteins / metabolism
  • CRISPR-Cas Systems*
  • Endonucleases / genetics*
  • Endonucleases / metabolism
  • Gene Library
  • Gene Silencing*
  • Gene Targeting / methods*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • K562 Cells
  • Machine Learning
  • RNA, Guide / genetics*
  • RNA, Guide / metabolism
  • Transfection

Substances

  • CRISPR-Associated Proteins
  • RNA, Guide
  • Endonucleases