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Recording RNA by CRISPR-Cas adaptation: a brief review
Sungchul Kim 1,* (Young Scientist Fellow / Principal Research Investigator), Gyeong-Seok Oh 1 (Postdoctoral researcher), Seongjin An 1,2 (Undergraduate student)
1Center for RNA Research, Institute for Basic Science, Seoul 08826, Korea,
2Department of Life Sciences, School of Life Sciences and Biotechnology, Korea University, Seoul, 02841, Korea
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. Non-self nucleic acid fragments are acquired as spacers between repeats in the host CRISPR array during a process called ‘adaptation’ 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 RNA-guided interference pathways. In some of 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 focused 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.
Abstract, Accepted Manuscript(in press) [Submitted on April 4, 2023, Accepted on April 4, 2023]
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