Flexible CRISPR library construction using parallel oligonucleotide retrieval

Nucleic Acids Res. 2017 Jun 20;45(11):e101. doi: 10.1093/nar/gkx181.

Abstract

CRISPR/Cas9-based gene knockout libraries have emerged as a powerful tool for functional screens. We present here a set of pre-designed human and mouse sgRNA sequences that are optimized for both high on-target potency and low off-target effect. To maximize the chance of target gene inactivation, sgRNAs were curated to target both 5΄ constitutive exons and exons that encode conserved protein domains. We describe here a robust and cost-effective method to construct multiple small sized CRISPR library from a single oligo pool generated by array synthesis using parallel oligonucleotide retrieval. Together, these resources provide a convenient means for individual labs to generate customized CRISPR libraries of variable size and coverage depth for functional genomics application.

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • Exons
  • Gene Knockout Techniques*
  • Gene Library
  • Humans
  • Mice
  • Oligonucleotides / genetics*

Substances

  • Oligonucleotides