US 2026/0015631 A1 · TYPE I-D CRISPR-ASSOCIATED TRANSPOSASE AND TYROSINE RECOMBINASE TRANSPOSON SYSTEMS

Abstract

The invention provides for systems and methods for inserting large polynucleotides into precise locations in a target polynucleotide. In one aspect, the systems comprise an engineered Type I-D/Tn7 CRISPR-associated transposase system (CAST) comprising a Tn7-like transposase linked to or otherwise capable of associating with a Type I-D CRISPR-Cas complex (Tn7-CAST I-D). In another aspect, the systems comprise a Tn7-like transposase comprising a modular target site selection protein, called TnsF, that may be engineered to reprogram the Tn7-like transposase to facilitate insertion at different sites in a target polynucleotide. In another aspect, the systems comprise a transposon system comprising a tyrosine recombinase which provides for scar-less insertion of large donor sequences into target polynucleotides. Also provided are methods for modifying target polynucleotides using the systems; polynucleotides encoding the systems; delivery systems for delivering the components of the systems; and cells and biological products modified by or modified to include the systems.

Claims150

Dependent claims

An engineered CRISPR-associated transposon (CAST) system comprising:

(a) one or more Type I-D Cas proteins;
(b) one or more CRISPR-associated Tn7 transposases or functional fragments thereof linked to or otherwise capable of associating with the one or more Type I-D Cas proteins;
and
(c) a guide molecule capable of forming a complex with the one or more Type I-D Cas proteins and directing sequence-specific binding of the complex to a target polynucleotide.

1 more element

Dependencies are inferred from the claim text.

Type i-d crispr-associated transposase and tyrosine recombinase transposon systems

Life of this patent

Type i-d crispr-associated transposase and tyrosine recombinase transposon systems

Life of this patent

Abstract

Claims150