⮝ Full datasets listing
PXD073781
PXD073781 is an original dataset announced via ProteomeXchange.
Dataset Summary
| Title | Multilayered regulation by RNA thermometers enables precise control of Cas9 expression in E. coli |
| Description | Cas9-based genome editing technologies can rapidly generate mutations to probe a diverse array of mutant genotypes. However, aberrant Cas9 nuclease translation and activity can occur despite the use of inducible promoters to control expression, leading to extensive cell death. This background killing caused by promoter leakiness severely limits the application of Cas9 for generating mutant libraries because of the potential for population skew. We demonstrate the utility of temperature sensitive RNA elements as a layer of post-transcriptional regulation to reduce the impact of promoter leak. We observe significant temperature-dependent increases in cell survival when certain RNA thermometers (RNATs) are placed upstream of the cas9 coding sequence. We also show that the most highly repressing RNAT, hsp17rep, significantly reduces population skew with a library of characterized guide RNAs in Escherichia coli. This strategy should be applicable to all bacterial Cas9-based methods and technologies. |
| HostingRepository | MassIVE |
| AnnounceDate | 2026-01-29 |
| AnnouncementXML | Submission_2026-01-29_07:19:47.533.xml |
| DigitalObjectIdentifier | |
| ReviewLevel | Non peer-reviewed dataset |
| DatasetOrigin | Original dataset |
| RepositorySupport | Unsupported dataset by repository |
| PrimarySubmitter | Richard J. Giannone |
| SpeciesList | scientific name: Escherichia coli; common name: E. coli; NCBI TaxID: 562; |
| ModificationList | iodoacetamide - site C; L-methionine sulfoxide |
| Instrument | Q Exactive Plus |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|---|---|---|
| 0 | 2026-01-29 06:54:59 | ID requested | |
| ⏵ 1 | 2026-01-29 07:19:48 | announced |
Publication List
| no publication |
Keyword List
| submitter keyword: E. coli, RNA, regulation, Cas9, CRISPR, DatasetType:Proteomics |
Contact List
| William G. Alexander | |
|---|---|
| contact affiliation | Oak Ridge National Laboratory |
| contact email | alexanderwg@ornl.gov |
| lab head | |
| Richard J. Giannone | |
| contact affiliation | Oak Ridge National Laboratory |
| contact email | giannonerj@ornl.gov |
| dataset submitter | |
Full Dataset Link List
| MassIVE dataset URI |
| Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://massive-ftp.ucsd.edu/v12/MSV000100641/ |
