FURA-PE3/AM

Prime editing is a CRISPR-derived genome editing method that uses a Cas9 H840A nickase fused to an engineered M-MLV reverse transcriptase and a prime-editing guide RNA (pegRNA), where the pegRNA specifies the genomic target through its spacer and encodes the desired edit through a primer-binding site and reverse-transcription template. The editor nicks one DNA strand, the exposed 3' DNA end hybridizes to the pegRNA primer-binding site, reverse transcription copies the edited sequence into a 3' flap, and cellular DNA repair resolves the intermediate to generate substitutions, small insertions, or small deletions without requiring a donor DNA template or a programmed double-strand break. The standard mammalian workflow designs candidate pegRNAs and, when using PE3 or PE3b, an additional nicking sgRNA; delivers prime-editor and guide components into cells; allows editing to occur; extracts genomic DNA; amplifies the target locus; and quantifies precise edits, indels, and byproducts by amp