Tandem repeat knockout utilizing the CRISPR/Cas9 system in human cells

Gene. 2016 May 15;582(2):122-7. doi: 10.1016/j.gene.2016.02.013. Epub 2016 Feb 9.

Abstract

Tandem repeats have been shown to cause human genetic diseases and contribute significantly to genome variation and instability. Although multi-sgRNAs mediated CRISPR/Cas9 system have used to generate regional deletions previously, in this study we explored a method of generating regional deletions of tandem repeats by taking advantage of the off-target effects of CRISPR/Cas9 in 293FT cells. Our results revealed that generation of large-fragment deletions of tandem repeats located in the MAGEL2 and XIST gene was possible. In summary, we have demonstrated that large-fragment deletions of tandem repeats can be achieved using a sgRNA-directed CRISPR/Cas9 system, facilitating the functional study of tandem repeats in future studies.

Keywords: CRISPR/Cas9; Off-target mutation; Regional deletions; Tandem repeats.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • CRISPR-Cas Systems / genetics*
  • Gene Knockout Techniques*
  • Genetic Vectors / metabolism
  • Genome, Human
  • HEK293 Cells
  • Humans
  • Molecular Sequence Data
  • RNA, Guide, CRISPR-Cas Systems / genetics
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / metabolism
  • Tandem Repeat Sequences / genetics*

Substances

  • RNA, Guide, CRISPR-Cas Systems
  • RNA, Long Noncoding
  • XIST non-coding RNA