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. Author manuscript; available in PMC: 2018 Feb 1.
Published in final edited form as: Cell Microbiol. 2016 Aug 25;19(2):10.1111/cmi.12651. doi: 10.1111/cmi.12651

Figure 4.

Figure 4

Complete Knockout of Selected Genes. A. Scheme of CRISPR/Cas9-mediated gene knockout using short-homology flanked resistance cassette. PCR primer positions are indicated by arrows. B. Agarose gel electrophoresis of PCR1 products for single gene knockout parasites. PCR1 was conducted on parental PruΔku80Δhxgprt wild type and the corresponding knockout parasite for the CDS of the gene of interest. PCR was conducted on clonal parasite lines, changes in PCR amplicon sizes were consistent with predicted sizes for wild type loci and those replaced by the resistance cassettes C. Agarose gel electrophoresis of PCR1 products for double knockout parasites. PCR was conducted across the CDS region of the gene of interest in a single gene knockout parental strain and the corresponding double knockouts.