期刊
CRISPR JOURNAL
卷 3, 期 5, 页码 398-408出版社
MARY ANN LIEBERT, INC
DOI: 10.1089/crispr.2020.0029
关键词
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资金
- 7th Framework Programme [610730, 613745]
- Horizon 2020 Marie SklodowskaCurie [642738]
- Engineering and Physical Sciences Research Council (EPSRC)
- Biotechnology and Biological Sciences Research Council (BBSRC) [BB/P020615/1, BB/M017982/1]
- School of Life Sciences (University of Warwick)
- EPSRC/BBSRC [BB/M017982/1]
- DGA-Dstl fellowship
- BBSRC [BB/P020615/1, BB/M017982/1] Funding Source: UKRI
CRISPR guide RNAs (gRNAs) can be programmed with relative ease to allow the genetic editing of nearly any DNA or RNA sequence. Here, we propose novel molecular architectures to achieve RNA-dependent modulation of CRISPR activity in response to specific RNA molecules. We designed and tested, in both living Escherichia coli cells and cell-free assays for rapid prototyping, cis-repressed RNA-interacting guide RNA (igRNA) that switch to their active state only upon interaction with small RNA fragments or long RNA transcripts, including pathogen-derived mRNAs of medical relevance such as the human immunodeficiency virus infectivity factor. The proposed CRISPR-igRNAs are fully customizable and easily adaptable to the majority if not all the available CRISPR-Cas variants to modulate a variety of genetic functions in response to specific cellular conditions, providing orthogonal activation and increased specificity. We thereby foresee a large scope of application for therapeutic, diagnostic, and biotech applications in both prokaryotic and eukaryotic systems.
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