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Rational gRNA desig...
Rational gRNA design based on transcription factor binding data
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- Bergenholm, David, 1987 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Dabirian, Yasaman, 1992 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Ferreira, Raphael, 1990 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Siewers, Verena, 1976 (författare)
- BioInnovation Institute (BII)
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- David, Florian, 1981 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Nielsen, Jens B, 1962 (författare)
- Chalmers tekniska högskola,Chalmers University of Technology
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(creator_code:org_t)
- 2021-07-27
- 2021
- Engelska.
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Ingår i: Synthetic Biology. - : Oxford University Press (OUP). - 2397-7000 .- 1939-7267. ; 6:1
- Relaterad länk:
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https://research.cha... (primary) (free)
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https://academic.oup...
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https://doi.org/10.1...
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https://research.cha...
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Abstract
Ämnesord
Stäng
- The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system has become a standard tool in many genome engineering endeavors. The endonuclease-deficient version of Cas9 (dCas9) is also a powerful programmable tool for gene regulation. In this study, we made use of Saccharomyces cerevisiae transcription factor (TF) binding data to obtain a better understanding of the interplay between TF binding and binding of dCas9 fused to an activator domain, VPR. More specifically, we targeted dCas9-VPR toward binding sites of Gcr1-Gcr2 and Tye7 present in several promoters of genes encoding enzymes engaged in the central carbon metabolism. From our data, we observed an upregulation of gene expression when dCas9-VPR was targeted next to a TF binding motif, whereas a downregulation or no change was observed when dCas9 was bound on a TF motif. This suggests a steric competition between dCas9 and the specific TF. Integrating TF binding data, therefore, proved to be useful for designing guide RNAs for CRISPR interference or CRISPR activation applications.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
- NATURVETENSKAP -- Biologi -- Bioinformatik och systembiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Bioinformatics and Systems Biology (hsv//eng)
Nyckelord
- glycolytic promoters
- transcription factor binding
- gRNA design
- Saccharomyces cerevisiae
- CRISPRa
Publikations- och innehållstyp
- art (ämneskategori)
- ref (ämneskategori)
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