Back to results
US 2025/0084390 A1Published application
Recruitment of dna polymerase for templated editing
AssigneePairwise Plants Services, Inc.
InventorsJingyi Nie, Aaron Hummel, Shai Joshua Lawit
Life of this patent
Estimating the term…
Abstract
This invention relates to recombinant nucleic constructs comprising a sequence-specific DNA binding protein, DNA-dependent DNA polymerase and a DNA encoded repair template, optionally a DNA endonuclease or wherein the sequence-specific DNA binding protein comprises DNA endonuclease activity, and methods of use thereof for modifying nucleic acids in cells and organisms.
Claims20
A method of modifying a target nucleic acid, the method comprising:
- contacting the target nucleic acid with:
- (a) a sequence-specific DNA binding protein that binds to a site on a target nucleic acid, wherein the sequence-specific DNA binding protein is from a CRISPR-Cas effector protein;
- (b) a DNA endonuclease having nickase or nuclease activity that introduces a single strand nick or a double strand break in the target nucleic acid;
- (c) an RNA:
- DNA hybrid guide comprising an RNA guide nucleic acid and a DNA encoded repair template, wherein the DNA encoded repair template is linked to the RNA guide nucleic acid, thereby guiding the DNA encoded repair template to the target nucleic acid;
- and
- (d) a DNA-dependent DNA polymerase that initiates strand synthesis copying genetic information from the DNA encoded repair template to the target nucleic acid, wherein the DNA-dependent DNA polymerase is recruited to the sequence-specific DNA binding protein, thereby modifying the target nucleic acid.
4 more elements
A first complex comprising:
- (a) a first sequence-specific DNA binding protein that is capable of binding to a first site on a target nucleic acid;
- and
- (b) a first DNA-dependent DNA polymerase.
A second complex comprising:
- (a) a second sequence-specific DNA binding protein that is capable of binding to a second site on a target nucleic acid;
- and
- (b) a DNA-encoded repair template.
Dependencies are inferred from the claim text.