Harnessing CRISPR-Cas adaptation for RNA recording and beyond

BMB Rep. 2024 Jan;57(1):40-49. doi: 10.5483/BMBRep.2023-0050.

Abstract

Prokaryotes encode clustered regularly interspaced short palindromic repeat (CRISPR) arrays and CRISPR-associated (Cas) genes as an adaptive immune machinery. CRISPR-Cas systems effectively protect hosts from the invasion of foreign enemies, such as bacteriophages and plasmids. During a process called 'adaptation', non-self-nucleic acid fragments are acquired as spacers between repeats in the host CRISPR array, to establish immunological memory. The highly conserved Cas1-Cas2 complexes function as molecular recorders to integrate spacers in a time course manner, which can subsequently be expressed as crRNAs complexed with Cas effector proteins for the RNAguided interference pathways. In some of the RNA-targeting type III systems, Cas1 proteins are fused with reverse transcriptase (RT), indicating that RT-Cas1-Cas2 complexes can acquire RNA transcripts for spacer acquisition. In this review, we summarize current studies that focus on the molecular structure and function of the RT-fused Cas1-Cas2 integrase, and its potential applications as a directional RNA-recording tool in cells. Furthermore, we highlight outstanding questions for RT-Cas1-Cas2 studies and future directions for RNA-recording CRISPR technologies. [BMB Reports 2024; 57(1): 40-49].

Publication types

  • Review
  • News

MeSH terms

  • Bacterial Proteins / metabolism
  • CRISPR-Cas Systems* / genetics
  • Integrases / genetics
  • Plasmids
  • RNA* / genetics

Substances

  • RNA
  • Bacterial Proteins
  • Integrases

Grants and funding

ACKNOWLEDGEMENTS This work was supported by IBS-R008-D1, Young Scientist Fellowship program of the Institute for Basic Science from the Ministry of Science and ICT of Korea.