Simple method for assembly of CRISPR synergistic activation mediator gRNA expression array

J Biotechnol. 2018 May 20:274:54-57. doi: 10.1016/j.jbiotec.2018.03.018. Epub 2018 Mar 26.

Abstract

When studying complex interconnected regulatory networks, effective methods for simultaneously manipulating multiple genes expression are paramount. Previously, we have developed a simple method for generation of an all-in-one CRISPR gRNA expression array. We here present a Golden Gate Assembly-based system of synergistic activation mediator (SAM) compatible CRISPR/dCas9 gRNA expression array for the simultaneous activation of multiple genes. Using this system, we demonstrated the simultaneous activation of the transcription factors, TWIST, SNAIL, SLUG, and ZEB1 a human breast cancer cell line.

Keywords: CRISPR; CRISPRa; Gene activation; Golden gate assembly; SAM.

MeSH terms

  • CRISPR-Cas Systems
  • Cell Line, Tumor
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation, Neoplastic
  • Gene Regulatory Networks*
  • Humans
  • Neoplasms / genetics
  • Oligonucleotide Array Sequence Analysis / methods*
  • RNA, Guide, CRISPR-Cas Systems / genetics*

Substances

  • RNA, Guide, CRISPR-Cas Systems