Barbier, I and Kusumawardhani, H and Chauhan, L and Harlapur, PV and Jolly, MK and Schaerli, Y (2023) Synthetic Gene Circuits Combining CRISPR Interference and CRISPR Activation in E. coli: Importance of Equal Guide RNA Binding Affinities to Avoid Context-Dependent Effects. In: ACS Synthetic Biology, 12 (10). pp. 3064-3071.
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Abstract
Gene expression control based on clustered regularly interspaced short palindromic repeats (CRISPR) has emerged as a powerful approach for constructing synthetic gene circuits. While the use of CRISPR interference (CRISPRi) is already well-established in prokaryotic circuits, CRISPR activation (CRISPRa) is less mature, and a combination of the two in the same circuits is only just emerging. Here, we report that combining CRISPRi with SoxS-based CRISPRa in Escherichia coli can lead to context-dependent effects due to different affinities in the formation of CRISPRa and CRISPRi complexes, resulting in loss of predictable behavior. We show that this effect can be avoided by using the same scaffold guide RNA structure for both complexes. © 2023 The Authors. Published by American Chemical Society.
| Item Type: | Journal Article |
|---|---|
| Publication: | ACS Synthetic Biology |
| Publisher: | American Chemical Society |
| Additional Information: | The copyright for this article belongs to authors. |
| Keywords: | arabinose; DNA; guide RNA; messenger RNA; small cytoplasmic RNA; synthetic DNA; RNA; synthetic DNA, Article; binding affinity; binding site; clustered regularly interspaced short palindromic repeat; clustered regularly interspaced short palindromic repeat activation; clustered regularly interspaced short palindromic repeat interference; competition; complex formation; controlled study; DNA binding; Escherichia coli; mathematical model; nonhuman; regulatory mechanism; RNA analysis; RNA binding; RNA structure; synthetic biology; clustered regularly interspaced short palindromic repeat; CRISPR Cas system; genetics; metabolism, Clustered Regularly Interspaced Short Palindromic Repeats; CRISPR-Cas Systems; Escherichia coli; Genes, Synthetic; RNA |
| Department/Centre: | Division of Interdisciplinary Sciences > Centre for Biosystems Science and Engineering |
| Date Deposited: | 10 Dec 2024 18:34 |
| Last Modified: | 10 Dec 2024 18:34 |
| URI: | http://eprints.iisc.ac.in/id/eprint/85397 |
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