Development and Characterization of a Modular CRISPR and RNA Aptamer Mediated Base Editing System

CRISPR J. 2021 Feb;4(1):58-68. doi: 10.1089/crispr.2020.0035.

Abstract

Conventional CRISPR approaches for precision genome editing rely on the introduction of DNA double-strand breaks (DSB) and activation of homology-directed repair (HDR), which is inherently genotoxic and inefficient in somatic cells. The development of base editing (BE) systems that edit a target base without requiring generation of DSB or HDR offers an alternative. Here, we describe a novel BE system called Pin-pointTM that recruits a DNA base-modifying enzyme through an RNA aptamer within the gRNA molecule. Pin-point is capable of efficiently modifying base pairs in the human genome with precision and low on-target indel formation. This system can potentially be applied for correcting pathogenic mutations, installing premature stop codons in pathological genes, and introducing other types of genetic changes for basic research and therapeutic development.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aptamers, Nucleotide*
  • Bacteria / genetics
  • Bacteria / metabolism
  • CRISPR-Cas Systems
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Exome Sequencing
  • Gene Editing*
  • Green Fluorescent Proteins / genetics
  • HEK293 Cells
  • Humans
  • INDEL Mutation
  • RNA Editing*
  • RNA, Guide, CRISPR-Cas Systems / genetics
  • Recombinational DNA Repair

Substances

  • Aptamers, Nucleotide
  • RNA, Guide, CRISPR-Cas Systems
  • Green Fluorescent Proteins