Sequence-specific inhibition of microRNA via CRISPR/CRISPRi system

Sci Rep. 2014 Feb 3;4:3943. doi: 10.1038/srep03943.

Abstract

Here, we report a convenient and efficient miRNA inhibition strategy employing the CRISPR system. Using specifically designed gRNAs, miRNA gene has been cut at a single site by Cas9, resulting in knockdown of the miRNA in murine cells. Using a modified CRISPR interference system (CRISPRi), inactive Cas9 can reversibly prevent the expression of both monocistronic miRNAs and polycistronic miRNA clusters. Furthermore, CRISPR/CRISPRi is also capable of suppressing genes in porcine cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Animals
  • Bacterial Proteins / metabolism
  • CRISPR-Associated Protein 9
  • Cell Line
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Endonucleases / metabolism
  • Gene Expression
  • Gene Targeting / methods*
  • Green Fluorescent Proteins / genetics
  • Mice
  • MicroRNAs / antagonists & inhibitors*
  • MicroRNAs / genetics
  • RNA, Guide / genetics
  • RNA, Small Interfering / antagonists & inhibitors*
  • RNA, Small Interfering / genetics
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Swine

Substances

  • Bacterial Proteins
  • MicroRNAs
  • RNA, Guide
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • CRISPR-Associated Protein 9
  • Cas9 protein, Francisella novicida
  • Endonucleases