Reprogrammed tracrRNAs enable repurposing of RNAs as crRNAs and sequence-specific RNA biosensors

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dc.contributor.authorLiu, Yang
dc.contributor.authorPinto, Filipe
dc.contributor.authorWan, Xinyi
dc.contributor.authorYang, Zhugen
dc.contributor.authorPeng, Shuguang
dc.contributor.authorLi, Mengxi
dc.contributor.authorCooper, Jonathan M.
dc.contributor.authorXie, Zhen
dc.contributor.authorFrench, Christopher E.
dc.contributor.authorWang, Baojun
dc.date.accessioned2022-04-27T10:56:14Z
dc.date.available2022-04-27T10:56:14Z
dc.date.embargo2022-04-27
dc.date.issued2022-04-11
dc.description.abstractIn type II CRISPR systems, the guide RNA (gRNA) comprises a CRISPR RNA (crRNA) and a hybridized trans-acting CRISPR RNA (tracrRNA), both being essential in guided DNA targeting functions. Although tracrRNAs are diverse in sequence and structure across type II CRISPR systems, the programmability of crRNA-tracrRNA hybridization for Cas9 is not fully understood. Here, we reveal the programmability of crRNA-tracrRNA hybridization for Streptococcus pyogenes Cas9, and in doing so, redefine the capabilities of Cas9 proteins and the sources of crRNAs, providing new biosensing applications for type II CRISPR systems. By reprogramming the crRNA-tracrRNA hybridized sequence, we show that engineered crRNA-tracrRNA interactions can not only enable the design of orthogonal cellular computing devices but also facilitate the hijacking of endogenous small RNAs/mRNAs as crRNAs. We subsequently describe how these re-engineered gRNA pairings can be implemented as RNA sensors, capable of monitoring the transcriptional activity of various environment-responsive genomic genes, or detecting SARS-CoV-2 RNA in vitro, as an Atypical gRNA-activated Transcription Halting Alarm (AGATHA) biosensor.en_UK
dc.identifier.citationLiu Y, Pinto F, Wan X, et al., (2022) Reprogrammed tracrRNAs enable repurposing of RNAs as crRNAs and sequence-specific RNA biosensors. Nature Communications, Volume 13, April 2022, Article number 1937en_UK
dc.identifier.issn2041-1723
dc.identifier.urihttps://doi.org/10.1038/s41467-022-29604-x
dc.identifier.urihttp://dspace.lib.cranfield.ac.uk/handle/1826/17815
dc.language.isoenen_UK
dc.publisherNature Publishing Groupen_UK
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjectCRISPR-Cas systemsen_UK
dc.subjectRNAen_UK
dc.subjectSynthetic biologyen_UK
dc.titleReprogrammed tracrRNAs enable repurposing of RNAs as crRNAs and sequence-specific RNA biosensorsen_UK
dc.typeArticleen_UK

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