1. Transient CRISPR-Cas9 system.
Transient CRISPR-Cas9 system. (A) Structures of CaCAS9 and sgRNA expression cassettes. The CaCAS9 gene was the Vyas codon-optimized version, expressed from the ENO1 promoter and flanked by a CYC1 terminator (3). The sgRNA was expressed under the SNR52 promoter and contained the ENO1 terminator (3). The guide sequence is 20 bp long and designed for each target sequence. (B) Schematic diagram of ADE2 deletion strategy. The CRISPR system produces double-strand breaks (DSB) at target sequences. The double-strand breaks can be resolved by homology-directed repair with the gene deletion templates, which have 80-bp arms homologous to the target gene, to create a homozygous deletion of ADE2.
Kyunghun Min et al. mSphere 2016; doi: /mSphere