17300668. METHODS AND COMPOSITIONS FOR EDITING NUCLEOTIDE SEQUENCES (Massachusetts Institute of Technology)

METHODS AND COMPOSITIONS FOR EDITING NUCLEOTIDE SEQUENCES

Organization Name

Massachusetts Institute of Technology

Inventor(s)

David R. Liu of Cambridge MA (US)

Andrew Vito Anzalone of Cambridge MA (US)

Max Walt Shen of Cambridge MA (US)

METHODS AND COMPOSITIONS FOR EDITING NUCLEOTIDE SEQUENCES

This abstract first appeared for US patent application 17300668 titled 'METHODS AND COMPOSITIONS FOR EDITING NUCLEOTIDE SEQUENCES

Original Abstract Submitted

The present disclosure provides new prime editor guide RNAs for prime editing, constructs for prime editing, and methods for using same. In addition, the present disclosure provides compositions and methods for conducting prime editing of a target DNA molecule (e.g., a genome) that enables the incorporation of a nucleotide change and/or targeted mutagenesis (e.g., insertion or deletion). The nucleotide change can include a single-nucleotide change (e.g., any transition or any transversion), an insertion of one or more nucleotides, or a deletion of one or more nucleotides. More in particular, the disclosure provides fusion proteins comprising nucleic acid programmable DNA binding proteins (napDNAbp) and a polymerase (e.g., reverse transcriptase), which is guided to a specific DNA sequence by a prime editor RNA (PEgRNA). The prime editor guide RNA comprises an extension arm that provides a DNA synthesis template sequence which encodes a single strand DNA flap, which is homologous to an endogenous DNA sequence, but which contains the desired one or more nucleotide changes and which, following synthesis by the polymerase (e.g., reverse transcriptase), becomes incorporated into the target DNA molecule.