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US 2023/0040216 A1Published application
Retrotransposons and use thereof
AssigneesThe Broad Institute, Inc., Massachusetts Institute of Technology
InventorsFeng Zhang, Michael Segel, Alim Ladha
Life of this patent
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Abstract
Systems and methods for targeted gene modification, targeted insertion, perturbation of gene transcripts, and nucleic acid editing. Novel nucleic acid targeting systems comprise components of CRISPR systems and non-LTR retrotransposon elements.
Claims75
An engineered or non-naturally occurring composition comprising:
- a. a site-specific nuclease polypeptide, or a polynucleotide comprising a coding sequence thereof;
- b. a non-LTR retrotransposon polypeptide connected to or otherwise capable of forming a complex with the site-specific nuclease polypeptide, or a polynucleotide comprising a coding sequence thereof;
- c. a guide molecule capable of forming a complex with the site-specific nuclease polypeptide and directing site-specific binding to a target sequence of a target polynucleotide;
- and
- d. a polynucleotide encoding a retrotransposon RNA, wherein the retrotransposon RNA comprises or encodes a donor polynucleotide.
2 more elements
An engineered or non-naturally occurring composition comprising:
- a. two site-specific nuclease polypeptides, or one or more polynucleotides comprising coding sequences thereof;
- b. two non-LTR retrotransposon polypeptides, each connected to or otherwise capable of forming a complex with one of the two site-specific nuclease polypeptides, or one or more polynucleotides comprising coding sequences thereof;
- c. two guide molecules, each capable of forming a complex with one of the site-specific nuclease polypeptides and directing site-specific binding to a target sequence of a target polynucleotide;
- and
- d. a polynucleotide encoding a retrotransposon RNA comprising or encoding a donor polynucleotide.
2 more elements
An engineered composition for non-native, targeted transposition of donor sequence into targeted nucleic acids, comprising:
- a. a fusion protein comprising a site-specific nuclease fused to the N-terminus of a non-LTR retrotransposon polypeptide, or a polynucleotide comprising a coding sequence thereof;
- and
- b. a donor construct comprising a donor polynucleotide sequence located between two binding elements capable of forming a complex with the non-LTR retrotransposon polypeptide.
Dependencies are inferred from the claim text.