CRISPR prime editing with ribonucleoprotein complexes in zebrafish and primary human cells

Nat Biotechnol. 2022 Feb;40(2):189-193. doi: 10.1038/s41587-021-00901-y. Epub 2021 Apr 29.

Abstract

Prime editors have been delivered using DNA or RNA vectors. Here we demonstrate prime editing with purified ribonucleoprotein complexes. We introduced somatic mutations in zebrafish embryos with frequencies as high as 30% and demonstrate germline transmission. We also observed unintended insertions, deletions and prime editing guide RNA (pegRNA) scaffold incorporations. In HEK293T and primary human T cells, prime editing with purified ribonucleoprotein complexes introduced desired edits with frequencies of up to 21 and 7.5%, respectively.

Publication types

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

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats* / genetics
  • Gene Editing
  • HEK293 Cells
  • Humans
  • RNA, Guide, Kinetoplastida / genetics
  • Ribonucleoproteins / genetics
  • Zebrafish* / genetics

Substances

  • RNA, Guide
  • Ribonucleoproteins