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OffScan: a universal and fast CRISPR off-target sites detection tool

BMC Genomics. 2020 Mar 5;21(Suppl 1):872. doi: 10.1186/s12864-019-6241-9.

Abstract

Background: The Type II clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) is a powerful genome editing technology, which is more and more popular in gene function analysis. In CRISPR/Cas, RNA guides Cas nuclease to the target site to perform DNA modification.

Results: The performance of CRISPR/Cas depends on well-designed single guide RNA (sgRNA). However, the off-target effect of sgRNA leads to undesired mutations in genome and limits the use of CRISPR/Cas. Here, we present OffScan, a universal and fast CRISPR off-target detection tool.

Conclusions: OffScan is not limited by the number of mismatches and allows custom protospacer-adjacent motif (PAM), which is the target site by Cas protein. Besides, OffScan adopts the FM-index, which efficiently improves query speed and reduce memory consumption.

Keywords: CRISPR/Cas; FM-index; Off-target; sgRNA.

MeSH terms

  • Algorithms
  • Animals
  • CRISPR-Cas Systems*
  • Caenorhabditis elegans / genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Computational Biology / methods*
  • Endonucleases / metabolism
  • Gene Editing / methods*
  • Humans
  • Mice
  • Mutation
  • RNA, Guide, CRISPR-Cas Systems / genetics*
  • Zebrafish / genetics

Substances

  • RNA, Guide, CRISPR-Cas Systems
  • Endonucleases