Chunlei Jiao         email
Helmholtz Institute for RNA-based Infection Research  

Supervisor:
Prof. Dr. Chase Beisel (Würzburg)
Promotion Committee:
Prof. Dr. Chase Beisel (Würzburg)
Prof. Dr. Cynthia Sharma (Würzburg)
Jun. Prof. Dr. Redmond Smyth (Würzburg)

 

 

 

Developing a programmable mRNA-derived crRNA(mcrRNA) based CRISPR-Cas9 technology

Although CRISPR-Cas9 system has been widely used for genome editing in various fields, like agriculture, industry and medicine, there is an increasing demand for precise time and spatial expression of CRISPR-Cas9 system to mitigate its off-targeting and cytotoxicity. Some switchable CRISPR-Cas9 systems have been developed to respond to external signals, like small molecules, light and temperature.

The typical native CRISPR-Cas system consists dual RNAs, TracrRNA and CrRNA. The repeat region in crRNA matchs with the antirepeat region in TracrRNA by base-pairing, although there are some nucleotides mismatching in the middle of repeat-antirepeat region. Some studies showed that SpyCas9 is tolerant to the nucleotides substitution (still keeping base-pairing) in the repeat-antirepeat region except for the part with nucleotides mismatching.

Based on the findings, my PhD project aims to develop a programmable mRNA-derived crRNA (mcrRNA) based CRISPR-Cas9 technology by tailoring TracrRNA to match any endogenous mRNA and produce mature mcrRNA to guide the Cas9 cleavage in the corresponding site in the genome