Weiting Zhang, PhD

Pronouns

She/Her/Hers

Rank

Research Fellow

Institution

MGH

BWH-MGH Title

Department

Medicine

Authors

Karl Petri, Weiting Zhang, Junyan Ma, Andrea Schmidts, Hyunho Lee, Joy E. Horng, Daniel Y. Kim, Ibrahim C. Kurt, Kendell Clement, Jonathan Y. Hsu, Luca Pinello, Marcela V. Maus, J. Keith Joung, Jing-Ruey Joanna Yeh

Prime editing with purified ribonucleoprotein complexes in zebrafish

WMSS 2022

In the first place, I would like to support women in science by participating in this event. Second, it would be a great honor for me to be selected by the organizers and encourage me to pursue science.

Our lab has been developing more efficient, precise, and easier-to-use gene editing tools for zebrafish research. With these tools, we and others will be able to generate zebrafish mutations mimicking human disease-causing mutations. The overarching goals are to use these zebrafish models for dissecting disease mechanisms and investigating treatment options.

Background

CRISPR technologies have brought a new era in gene editing. Prime editing is a novel method of genome editing on CRISPR systems. It employs a pegRNA and a prime editing protein consisting of a Cas9 nickase fused to an engineered M-MLV reverse transcriptase. Compared to previous CRISPR technologies, prime editing has higher precision in creating any user-defined small DNA modifications.

Methods

We overexpressed and purified prime editing protein from Escherichia coli. Ribonucleoprotein complexes of prime editors were injected into zebrafish embryos. Next gene sequencing was used to identify editing efficiency.

Results

Prime editors have been delivered using DNA or RNA vectors. We and our collaborators demonstrate prime editing with purified ribonucleoprotein complexes in zebrafish and primary human cells. Our work has been published in Nature Biotechnology in 2021.

Conclusions

We have developed a platform for researchers to apply prime editing to zebrafish which enable scientists to create useful animal models mimicking human pathogenic mutations to uncover mechanisms of disease and potential therapeutic approaches.