Multiplexed CRISPR/Cas9 Genome Editing and Gene Regulation Using Csy4 in Saccharomyces cerevisiae

Ferreira, Raphael,1990Chalmers tekniska högskola,Chalmers University of Technology (författare)

Clustered regularly interspaced short palindromic repeats (CRISPR) technology has greatly accelerated the field of strain engineering. However, insufficient efforts have been made toward developing robust multiplexing tools in Saccharomyces cerevisiae. Here, we exploit the RNA processing capacity of the bacterial endoribonuclease Csy4 from Pseudomonas aeruginosa, to generate multiple gRNAs from a single transcript for genome editing and gene interference applications in S. cerevisiae. In regards to genome editing, we performed a quadruple deletion of FAA1, FAA4, POX1 and TES1 reaching 96% efficiency out of 24 colonies tested. Then, we used this system to efficiently transcriptionally regulate the three genes, OLE1, HMG1 and ACS1. Thus, we demonstrate that multiplexed genome editing and gene regulation can be performed in a fast and effective manner using Csy4.

Skrekas, Christos,1990Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)skrekas (författare)
Nielsen, Jens B,1962Danmarks Tekniske Universitet,Technical University of Denmark,Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)nielsenj (författare)
David, Florian,1981Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)davidfl (författare)
Chalmers tekniska högskolaDanmarks Tekniske Universitet (creator_code:org_t)

Ingår i:ACS Synthetic Biology: American Chemical Society (ACS)7:1, s. 10-152161-5063

Av författaren/redakt...: Ferreira, Raphae ...; Skrekas, Christo ...; Nielsen, Jens B, ...; David, Florian, ...

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