C. albicans CRISPR-Cas9 Plasmid Toolkit

The plasmid backbone used in this study was adapted from the C. albicans-optimized CRISPR-Cas9 plasmid (also known as pRS252) used in our previous study (33 (link)), containing the NEUT5L homology site and CAS9 (79 (link)). To create a sgRNA cloning locus in this plasmid, the SNR52 promoter, SapI cloning locus, and sgRNA tail were synthesized in vitro as gBlocks gene fragments from Integrated DNA Technologies (IDT) and were cloned into the CRISPR-Cas9 plasmid (pRS252) at the NgoMIV restriction enzyme site, using Gibson assembly, as previously described (33 (link), 79 (link)). We have made the relevant CRISPRi (dCas9 and dCas9-Mxi1) plasmids available via Addgene (reference numbers 122377, 122378, 122379, and 122380).

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Wensing L., Sharma J., Uthayakumar D., Proteau Y., Chavez A, & Shapiro R.S. (2019). A CRISPR Interference Platform for Efficient Genetic Repression in Candida albicans. mSphere, 4(1), e00002-19.

Publication 2019

gBlocks gene fragments

Backbone Crispr Gene Mxi1 Plasmids Restriction enzyme Tail

Corresponding Organization :

Other organizations : University of Guelph, Columbia University

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Variable analysis

independent variables

The plasmid backbone used in this study was adapted from the C. albicans-optimized CRISPR-Cas9 plasmid (also known as pRS252) used in our previous study (33 (link)).
The SNR52 promoter, SapI cloning locus, and sgRNA tail were synthesized in vitro as gBlocks gene fragments and were cloned into the CRISPR-Cas9 plasmid (pRS252) at the NgoMIV restriction enzyme site, using Gibson assembly.

dependent variables

Not explicitly mentioned.

control variables

The NEUT5L homology site and CAS9 were contained in the plasmid backbone used in this study.
The relevant CRISPRi (dCas9 and dCas9-Mxi1) plasmids were made available via Addgene (reference numbers 122377, 122378, 122379, and 122380).

controls

No positive or negative controls were explicitly mentioned.

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