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. 2020 Sep 3;10:14605. doi: 10.1038/s41598-020-71648-w

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Design of the CRISPRa/i toolkit for transcriptional modulation. (a) The CRISPRa/i technology utilizes a catalytically inactive Cas9 (dCas9) to modulate the expression of genes targeted by an sgRNA. This can be further increased by fusing activation or repressor domains to dCas9. (b) Schematic representation of the CRISPRa/i components included in the CRISPRa/i plasmids and of the sgRNA expression cassette, present in all the plasmids. (c) The target-specific sgRNA from a double stranded oligonucleotide (ds oligo) was inserted into the CRISPRa/i plasmid through in vivo homologous recombination, allowing direct phenotypic characterization of strains with altered gene expression.