Ways of improving precise knock-in by genome-editing technologies

Hum Genet. 2019 Jan;138(1):1-19. doi: 10.1007/s00439-018-1953-5. Epub 2018 Nov 2.


Despite the recent discover of genome-editing methods, today we can say these approaches have firmly entered our life. Two approaches-knocking out malfunctioning gene allele or correcting the mutation with precise knock-in-can be used in hereditary monogenic diseases treatment. The latter approach is relatively ineffective. Modern data about the ways of repair of double-strand DNA breaks formed by nucleases are presented in this review. The main part of the review is devoted to the ways of increasing precise and effective knock-in: inhibition of non-homologous end joining and stimulation of homology-directed repair key factors, use of small molecules with unknown mechanism of action, cell-cycle synchronization and cell-cycle-dependent activity of Cas9, donor molecule design, selection, alternative methods for insertion and other approaches.

Publication types

  • Review

MeSH terms

  • CRISPR-Cas Systems*
  • Gene Editing / methods*
  • Gene Knock-In Techniques / methods*
  • Genetic Engineering / methods*
  • Humans