English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
Add to Basket
  Local TagsRelease HistoryDetailsSummary

Released
Journal Article

Selective Enrichment of Slow-Growing Bacteria in a Metabolism-Wide CRISPRi Library with a TIMER Protein

MPS-Authors
/persons/resource/persons254138

Beuter,  D.
Emmy Noether Research Group Dynamic Control of Metabolic Networks, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

Gomes,  J.
Max Planck Research Group Prokaryotic small RNA Biology, Alumni, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

/persons/resource/persons254626

Randau,  L.
Max Planck Research Group Prokaryotic small RNA Biology, Alumni, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

/persons/resource/persons263600

Diaz-Pascual,  Francisco
Max Planck Research Group Bacterial Biofilms, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

/persons/resource/persons254232

Drescher,  K.
Max Planck Research Group Bacterial Biofilms, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

/persons/resource/persons254499

Link,  H.
Emmy Noether Research Group Dynamic Control of Metabolic Networks, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

External Resource
No external resources are shared
Fulltext (restricted access)
There are currently no full texts shared for your IP range.
Fulltext (public)
There are no public fulltexts stored in PuRe
Supplementary Material (public)
There is no public supplementary material available
Citation

Beuter, D., Gomes, J., Randau, L., Diaz-Pascual, F., Drescher, K., & Link, H. (2018). Selective Enrichment of Slow-Growing Bacteria in a Metabolism-Wide CRISPRi Library with a TIMER Protein. ACS SYNTHETIC BIOLOGY, 7(12), 2775-2782. doi:10.1021/acssynbio.8b00379.


Cite as: https://hdl.handle.net/21.11116/0000-0004-465E-5
Abstract
Construction of pooled genetic variant libraries has become very fast and versatile. The current limitation of this technique is to select cells with a desired phenotype from very large libraries. Especially cells with poor fitness and slow growth are difficult to select because they are rapidly outcompeted by fitter cells. Here, we demonstrate selective and high-throughput enrichment of slow-growing strains using a fluorescent TIMER protein and flow cytometry. As a proof of principle, we created a metabolism-wide CRISPR interference library for Escherichia coli and enriched targets that interfere with amino acid metabolism. After enrichment of slow-growing cells, the CRISPRi library consisted almost entirely of targets that block amino acid biosynthesis. These results provide general guidelines for how to enrich slow-growing strains from a large pool of genetic variants, with applications in genetic screens, metabolic engineering, and synthetic biology.