UMass Chan Medical School Faculty Publications

Title

Deletion and replacement of long genomic sequences using prime editing

UMMS Affiliation

RNA Therapeutics Institute; Program in Bioinformatics and Integrative Biology; Li Weibo Institute for Rare Diseases Research

Publication Date

2021-10-14

Document Type

Article

Disciplines

Bioinformatics | Genetics and Genomics

Abstract

Genomic insertions, duplications and insertion/deletions (indels), which account for ~14% of human pathogenic mutations, cannot be accurately or efficiently corrected by current gene-editing methods, especially those that involve larger alterations (>100 base pairs (bp)). Here, we optimize prime editing (PE) tools for creating precise genomic deletions and direct the replacement of a genomic fragment ranging from ~1 kilobases (kb) to ~10 kb with a desired sequence (up to 60 bp) in the absence of an exogenous DNA template. By conjugating Cas9 nuclease to reverse transcriptase (PE-Cas9) and combining it with two PE guide RNAs (pegRNAs) targeting complementary DNA strands, we achieve precise and specific deletion and repair of target sequences via using this PE-Cas9-based deletion and repair (PEDAR) method. PEDAR outperformed other genome-editing methods in a reporter system and at endogenous loci, efficiently creating large and precise genomic alterations. In a mouse model of tyrosinemia, PEDAR removed a 1.38-kb pathogenic insertion within the Fah gene and precisely repaired the deletion junction to restore FAH expression in liver.

Keywords

Genetic engineering, Targeted gene repair

DOI of Published Version

10.1038/s41587-021-01026-y

Source

Jiang T, Zhang XO, Weng Z, Xue W. Deletion and replacement of long genomic sequences using prime editing. Nat Biotechnol. 2021 Oct 14. doi: 10.1038/s41587-021-01026-y. Epub ahead of print. PMID: 34650270. Link to article on publisher's site

Related Resources

Link to Article in PubMed

Journal/Book/Conference Title

Nature biotechnology

PubMed ID

34650270

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