Engineered RNA-Interacting CRISPR Guide RNAs for Genetic Sensing and Diagnostics
| Autor: | Charlotte M Quinn, John N. Duncan, Marko Storch, William Rostain, Roberto Galizi, Manish Kushwaha, Alfonso Jaramillo |
|---|---|
| Přispěvatelé: | Keele University [Keele], Imperial College London, University of Warwick [Coventry], Liverpool School of Tropical Medicine (LSTM), MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), This work was supported by the 7th Framework Programme (grant numbers 610730 [EVOPROG], 613745 [PROMYS]), the Horizon 2020 Marie SklodowskaCurie (grant number 642738 [MetaRNA]), the Engineering and Physical Sciences Research Council (EPSRC) and the Biotechnology and Biological Sciences Research Council (BBSRC, grant numbers BB/P020615/1 [EVO-ENGINE] and BB/M017982/1 [WISB Centre]), and the School of Life Sciences (University of Warwick, startup allocation) to A.J. Funding for open access charge: EPSRC/BBSRC (BB/M017982/1) to A.J. W.R. was supported by a DGA-Dstl fellowship., European Project: 610730,EC:FP7:ICT,FP7-ICT-2013-10,EVOPROG(2013), European Project: 613745,EC:FP7:KBBE,FP7-KBBE-2013-7-single-stage,PROMYS(2013), European Project: 642738,H2020,H2020-MSCA-ITN-2014,MetaRNA(2015), University of Liverpool, UK Research & Innovation (UKRI)Engineering & Physical Sciences Research Council (EPSRC), UK Research & Innovation (UKRI)Biotechnology and Biological Sciences Research Council (BBSRC)BB/P020615/1BB/M017982/1, School of Life Sciences (University of Warwick), UK Research & Innovation (UKRI) |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Small RNA
Transcription Genetic RNA-Interacting [SDV]Life Sciences [q-bio] Human immunodeficiency virus (HIV) Computational biology Biosensing Techniques Biology medicine.disease_cause Q1 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine qu_58.7 CRISPR-Associated Protein 9 Genetics medicine Escherichia coli vif Gene Products Human Immunodeficiency Virus CRISPR Clustered Regularly Interspaced Short Palindromic Repeats Active state Guide RNA RNA Messenger DNA Cleavage 030304 developmental biology 0303 health sciences [SDV.GEN]Life Sciences [q-bio]/Genetics Cell-Free System RNA 3. Good health QR qw_51 chemistry Genetic Sensing CRISPR-Cas Systems qu_470 Genetic Engineering 030217 neurology & neurosurgery DNA Biotechnology RNA Guide Kinetoplastida |
| Zdroj: | The Crispr Journal The Crispr Journal, 2020, 3 (5), pp.398-408. ⟨10.1089/crispr.2020.0029⟩ The CRISPR Journal The CRISPR Journal, 2020, 3 (5), pp.398-408. ⟨10.1089/crispr.2020.0029⟩ |
| ISSN: | 2573-1599 2573-1602 |
| Popis: | International audience; 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|