Improved prime editors enable pathogenic allele correction and cancer modelling in adult mice

• DOI: 10.1038/s41467-021-22295-w
• Publication Date: 2021-04-09T10:04:15Z
• Authors: Pengpeng Liu; Shun-Qing Liang; Chunwei Zheng; Esther Mintzer; Yan G. Zhao; Karthikeyan Ponni...; Aamir Mir; Erik J. Sontheimer; Guangping Gao; Terence R. Flotte; Scot A. Wolfe; Wen Xue
• Keywords: Prime editors; CRISPR-Cas9 genome editing; 0301 basic medicine; 570; Carcinogenesis; Science; 610; Bioengineering; Therapeutics; and Tissue Engineering; Transfection; Article; Mice; 03 medical and health sciences; Gene therapy; Nucleic Acids; Neoplasms; genome editing; Animals; Humans; Molecular Biology; Cancer genetics; Alleles; Cancer Biology; Gene Editing; Nucleotides; Q; Molecular; 500; Disease Modeling; Genetics and Genomics; Dependovirus; and Nucleosides; 3. Good health; Disease Models, Animal; HEK293 Cells; Cellular; CRISPR-Cas Systems; HeLa Cells

Abstract Prime editors (PEs) mediate genome modification without utilizing double-stranded DNA breaks or exogenous donor as a template. PEs facilitate nucleotide substitutions local insertions deletions within the based on template sequence encoded prime editing guide RNA (pegRNA). However, efficacy of in adult mice has not been established. Here we report an NLS-optimized SpCas9-based editor that improves efficiency both fluorescent reporter cells and at endogenous loci cultured cell lines. Using this system, could also seed tumor formation through somatic mouse. Finally, successfully utilize dual adeno-associated virus (AAVs) for delivery split-intein demonstrate system enables correction pathogenic mutation mouse liver. Our findings further establish broad potential technology directed installation modifications vivo, with important implications disease modeling correction.