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.
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.
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.
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.