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Effective CRISPRa-Mediated Control of Gene Expression in Bacteria Must Overcome Strict Target Site Requirements
- Source :
- Nature Communications, Vol 11, Iss 1, Pp 1-11 (2020), Nature Communications
- Publication Year :
- 2019
- Publisher :
- Cold Spring Harbor Laboratory, 2019.
-
Abstract
- In bacterial systems, CRISPR-Cas transcriptional activation (CRISPRa) has the potential to dramatically expand our ability to regulate gene expression, but we lack predictive rules for designing effective gRNA target sites. Here, we identify multiple features of bacterial promoters that impose stringent requirements on CRISPRa target sites. Notably, we observe narrow, 2–4 base windows of effective sites with a periodicity corresponding to one helical turn of DNA, spanning ~40 bases and centered ~80 bases upstream of the TSS. However, we also identify two features suggesting the potential for broad scope: CRISPRa is effective at a broad range of σ70-family promoters, and an expanded PAM dCas9 allows the activation of promoters that cannot be activated by S. pyogenes dCas9. These results provide a roadmap for future engineering efforts to further expand and generalize the scope of bacterial CRISPRa.<br />The use of CRISPRa in bacteria lacks predictive rules for identifying effective gRNA target sites. Here the authors define features of bacterial promoters that impose stringent requirements on effective sites.
- Subjects :
- Transcriptional Activation
0301 basic medicine
Computer science
Science
CRISPR-Associated Proteins
General Physics and Astronomy
Computational biology
Article
General Biochemistry, Genetics and Molecular Biology
Bacterial genetics
03 medical and health sciences
chemistry.chemical_compound
Gene expression
Escherichia coli
Clustered Regularly Interspaced Short Palindromic Repeats
Guide RNA
Promoter Regions, Genetic
Control (linguistics)
lcsh:Science
Gene
Synthetic biology
030304 developmental biology
Regulation of gene expression
0303 health sciences
Multidisciplinary
Bacteria
030102 biochemistry & molecular biology
Scope (project management)
biology
030306 microbiology
Escherichia coli Proteins
Promoter
Gene Expression Regulation, Bacterial
General Chemistry
biology.organism_classification
030104 developmental biology
Genetic circuit engineering
chemistry
Target site
Genes, Bacterial
Genetic engineering
Trans-Activators
lcsh:Q
CRISPR-Cas Systems
DNA
RNA, Guide, Kinetoplastida
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Nature Communications, Vol 11, Iss 1, Pp 1-11 (2020), Nature Communications
- Accession number :
- edsair.doi.dedup.....d8735a45384009aa072b4490f13bc42b
- Full Text :
- https://doi.org/10.1101/770891