Methods for CRISPR-Cas as Ribonucleoprotein Complex Delivery In Vivo

Mol Biotechnol. 2023 Feb;65(2):181-195. doi: 10.1007/s12033-022-00479-z. Epub 2022 Mar 24.

Abstract

The efficient delivery of CRISPR-Cas components is still a key and unsolved problem. CRISPR-Cas delivery in the form of a Cas protein+sgRNA (ribonucleoprotein complex, RNP complex), has proven to be extremely effective, since it allows to increase on-target activity, while reducing nonspecific activity. The key point for in vivo genome editing is the direct delivery of artificial nucleases and donor DNA molecules into the somatic cells of an adult organism. At the same time, control of the dose of artificial nucleases is impossible, which affects the efficiency of genome editing in the affected cells. Poor delivery efficiency and low editing efficacy reduce the overall potency of the in vivo genome editing process. Here we review how this problem is currently being solved in scientific works and what types of in vivo delivery methods of Cas9/sgRNA RNPs have been developed.

Keywords: CRISPR; Cas9; In vivo; Nanoparticles; RNP; Ribonucleoprotein complex.

Publication types

  • Review

MeSH terms

  • CRISPR-Cas Systems*
  • Gene Editing* / methods
  • Ribonucleoproteins / genetics
  • Ribonucleoproteins / metabolism

Substances

  • Ribonucleoproteins