WEHI PhD Completion Seminar hosted by Professor Marco Herold

Wei Jin

PhD Student – Herold Laboratory, Blood Cells and Blood Cancer division, WEHI

From Cas9 to Cas12a: An advanced CRISPR system to identify cancer drivers

Davis Auditorium

Join via SLIDO enter code #WEHIphdcompletion

Including Q&A session

Followed by refreshments in Tapestry Area

CRISPR technology has revolutionized biomedical research, by allowing fast and easy modelling and characterisation of cancer. Most CRISPR applications to date have relied on traditional Cas9 methodologies. To model more complex diseases, additional CRISPR platforms that can complement Cas9 mediated genome engineering are required.

During my PhD I characterised a mouse model that constitutively expresses enhanced Acidaminococcus sp.Cas12a (enAsCas12a) linked to an mCherry fluorescent reporter. Using primary and transformed cells from enAsCas12a mice, we demonstrate efficient single and multiplexed gene-editing in vitro, using. We further demonstrate successful in vivo gene-editing, using normal and cancer-prone enAsCas12a stem cells to reconstitute the haematopoietic system of wild-type mice. In addition, we developed compact, genome-wide Cas12a knockout libraries and demonstrated the utility across multiple screens in vitro and in vivo. Finally, we achieve CRISPR multiplexing via simultaneous gene knockout (via Cas12a) and activation (via dCas9-SAM) in different cell types.

Collectively, our enAsCas12a mouse model and accompanying crRNA expression libraries enhance genome engineering capabilities and complement current CRISPR technologies.

All welcome!