Editing of ccr5 receptor gene to protect against hiv infection
AssigneePresident and Fellows of Harvard College
InventorsJuan Pablo Maianti, David R. Liu
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Abstract
Provided herein are systems, compositions, and methods of introducing protective and/or loss-of-function variants of CCR5 and CCR2. Variants may be introduced using a CRISPR/Cas9-based nucleobase editor or other guide nucleotide sequence-programmable DNA binding protein domain-based fusion protein described herein. Further provided herein are compositions and methods of preventing and treating conditions related to HIV infection and progression as well as to AIDS.
Claims126
A method of editing a polynucleotide encoding a C-C chemokine receptor type five (CCR5) protein, the method comprising contacting the CCR5-encoding polynucleotide with:
- (i) a fusion protein comprising:
- (a) a guide nucleotide sequence-programmable DNA binding protein domain;
- and (b) a cytosine deaminase domain;
- and
- (ii) a guide nucleotide sequence targeting the fusion protein of (i) to a target cytosine (C) base in the CCR5-encoding polynucleotide;
- wherein the contacting results in the deamination of the target C base by the fusion protein, resulting in a cytosine-guanine (C:G) to thymine-adenine pair (T:A) change in the CCR5-encoding polynucleotide.
A method of editing a polynucleotide encoding a C-C chemokine receptor type 2 (CCR2) protein, the method comprising contacting the CCR2-encoding polynucleotide with:
- (i) a fusion protein comprising:
- (a) a guide nucleotide sequence-programmable DNA binding protein domain;
- and (b) a cytosine deaminase domain, and
- (ii) a guide nucleotide sequence targeting the fusion protein of (i) to a target cytosine (C) base in the CCR2-encoding polynucleotide,
- wherein the contacting results in the deamination of the target C base by the fusion protein, resulting in a cytosine-guanine (C:G) to thymine-adenine pair (T:A) change in the CCR2-encoding polynucleotide.
A method of editing a polynucleotide encoding a C-C chemokine receptor type five (CCR2) protein, the method comprising contacting the CCR2-encoding polynucleotide with:
- (i) a fusion protein comprising:
- (a) a guide nucleotide sequence-programmable DNA binding protein domain;
- and (b) a cytosine deaminase domain;
- and
- (ii) a guide nucleotide sequence targeting the fusion protein of (i) to a target cytosine (C) base in the CCR2-encoding polynucleotide;
- wherein the contacting results in the deamination of the target C base by the fusion protein, resulting in a cytosine-guanine (C:G) to thymine-adenine pair (T:A) change in the CCR2-encoding polynucleotide.
A composition comprising:
- (i) a fusion protein comprising:
- (a) a guide nucleotide sequence-programmable DNA binding protein domain;
- and (b) a cytosine deaminase domain;
- and
- (ii) a guide nucleotide sequence targeting the fusion protein of (i) to a polynucleotide encoding a C-C chemokine receptor type five (CCR5) protein.
A composition comprising:
- (i) a fusion protein comprising:
- (a) a guide nucleotide sequence-programmable DNA binding protein domain;
- and (b) a cytosine deaminase domain;
- and
- (ii) a guide nucleotide sequence targeting the fusion protein of (i) to a polynucleotide encoding a C-C chemokine receptor type two (CCR2) protein.
A composition comprising:
- (i) a fusion protein comprising:
- (a) a guide nucleotide sequence-programmable DNA binding protein domain;
- and (b) a cytosine deaminase domain;
- (ii) a guide nucleotide sequence targeting the fusion protein of (i) to a polynucleotide encoding a C-C chemokine receptor type five (CCR5) protein;
- and
- (iii) a guide nucleotide sequence targeting the fusion protein of (i) to a polynucleotide encoding a C-C chemokine receptor type 2 (CCR2) protein.
The method of claim C2, wherein the cell is selected from the group consisting of:
- macrophage, dendritic cell, memory T cell, endothelial cell, epithelial cell, vascular smooth muscle cell, fibroblast, microglia, neuron, and astrocyte.
Dependencies are inferred from the claim text.