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- W3207691330 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. Prime editing is expanded to deletions and replacements of genomic sequences of up to 10 kb." @default.
- W3207691330 created "2021-10-25" @default.
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- W3207691330 date "2021-10-14" @default.
- W3207691330 modified "2023-10-18" @default.
- W3207691330 title "Deletion and replacement of long genomic sequences using prime editing" @default.
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- W3207691330 doi "https://doi.org/10.1038/s41587-021-01026-y" @default.
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