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dc.contributor.authorHuang, Hsin-Ho
dc.contributor.authorBellato, Massimo
dc.contributor.authorQian, Yili
dc.contributor.authorCárdenas, Pablo
dc.contributor.authorPasotti, Lorenzo
dc.contributor.authorMagni, Paolo
dc.contributor.authorDel Vecchio, Domitilla
dc.date.accessioned2021-12-17T17:40:26Z
dc.date.available2021-12-17T17:40:26Z
dc.date.issued2021
dc.identifier.urihttps://hdl.handle.net/1721.1/138546
dc.description.abstractCRISPRi-mediated gene regulation allows simultaneous control of many genes. However, highly specific sgRNA-promoter binding is, alone, insufficient to achieve independent transcriptional regulation of multiple targets. Indeed, due to competition for dCas9, the repression ability of one sgRNA changes significantly when another sgRNA becomes expressed. To solve this problem and decouple sgRNA-mediated regulatory paths, we create a dCas9 concentration regulator that implements negative feedback on dCas9 level. This allows any sgRNA to maintain an approximately constant dose-response curve, independent of other sgRNAs. We demonstrate the regulator performance on both single-stage and layered CRISPRi-based genetic circuits, zeroing competition effects of up to 15-fold changes in circuit I/O response encountered without the dCas9 regulator. The dCas9 regulator decouples sgRNA-mediated regulatory paths, enabling concurrent and independent regulation of multiple genes. This allows predictable composition of CRISPRi-based genetic modules, which is essential in the design of larger scale synthetic genetic circuits.en_US
dc.language.isoen
dc.publisherSpringer Science and Business Media LLCen_US
dc.relation.isversionof10.1038/S41467-021-21772-6en_US
dc.rightsCreative Commons Attribution 4.0 International licenseen_US
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en_US
dc.sourceNatureen_US
dc.titledCas9 regulator to neutralize competition in CRISPRi circuitsen_US
dc.typeArticleen_US
dc.identifier.citationHuang, Hsin-Ho, Bellato, Massimo, Qian, Yili, Cárdenas, Pablo, Pasotti, Lorenzo et al. 2021. "dCas9 regulator to neutralize competition in CRISPRi circuits." Nature Communications, 12 (1).
dc.contributor.departmentMassachusetts Institute of Technology. Department of Mechanical Engineering
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biological Engineering
dc.contributor.departmentMassachusetts Institute of Technology. Synthetic Biology Center
dc.relation.journalNature Communicationsen_US
dc.eprint.versionFinal published versionen_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dc.date.updated2021-12-17T17:38:33Z
dspace.orderedauthorsHuang, H-H; Bellato, M; Qian, Y; Cárdenas, P; Pasotti, L; Magni, P; Del Vecchio, Den_US
dspace.date.submission2021-12-17T17:38:34Z
mit.journal.volume12en_US
mit.journal.issue1en_US
mit.licensePUBLISHER_CC
mit.metadata.statusAuthority Work and Publication Information Neededen_US


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