In certain aspects, the present invention provides methods for inducing a stable gene modification of a target nucleic acid via homologous recombination in a primary cell, such as a primary blood cell and/or a primary mesenchymal cell. In certain other aspects, the present invention provides methods for enriching a population of genetically modified primary cells having targeted integration at a target nucleic acid. The methods of the present invention rely on the introduction of a DNA nuclease such as a Cas polypeptide and a homologous donor adeno-associated viral (AAV) vector into the primary cell to mediate targeted integration of the target nucleic acid. Also provided herein are methods for preventing or treating a disease in a subject in need thereof by administering to the subject any of the genetically modified primary cells or pharmaceutical compositions described herein to prevent the disease or ameliorate one or more symptoms of the disease.

The present invention provides compositions and methods for transducing cells (e.g. T cells or immune cells). Also provided herein are methods of treating a disease in a subject in need thereof.

Certain aspects of the present invention relate to modified proteins e.g. oxygen-carrying proteins, comprising at least one modification for homogenous conjugation of one or more polymeric moieties e.g. polyethylene glycol or derivatives thereof.

The present invention relates to a genetically modified hematopoietic stem cell (HSC) comprising, in at least one α-globin gene comprised in the genome thereof, at least one transgene encoding a functional β-like globin protein, the said transgene being placed under the control of the endogenous promoter of the said at least one α-globin gene.

The present invention relates to a genetically modified hematopoietic stem cell (HSC) comprising, in at least one α-globin gene comprised in the genome thereof, at least one transgene encoding a functional β-like globin protein, the said transgene being placed under the control of the endogenous promoter of the said at least one α-globin gene.

Provided herein are ribonucleoprotein (RNP) complexes comprising a DNA-targeting endonuclease Cas (CRISPR-associated) protein and a guide RNA (gRNA) that that targets and hybridizes to the β-Globin gene. In one embodiment, the Cas protein is Cas9 and the gRNA comprises the sequence of SEQ ID NO: 1. In one embodiment, the Cas protein is Cas12a and the gRNA comprises the sequence of SEQ ID NO: 3.

Provided herein are ribonucleoprotein (RNP) complexes comprising a DNA-targeting endonuclease Cas (CRISPR-associated) protein and a guide RNA (gRNA) that that targets and hybridizes to the β-Globin gene. In one embodiment, the Cas protein is Cas9 and the gRNA comprises the sequence of SEQ ID NO: 1. In one embodiment, the Cas protein is Cas12a and the gRNA comprises the sequence of SEQ ID NO: 3.

Compositions that include a globin, such as hemoglobin, in a relaxed state are described. Globin molecules in a relaxed state (R state) have a higher binding affinity for carbon monoxide and oxygen than globin molecules in a tense state (T state). Hemoglobin in a relaxed state can be, for example, hemoglobin that is substantially free of 2,3-diphosphoglycerate or hemoglobin that includes a β-Cys93 that is covalently modified to inhibit one or both salt bridges between β-Asp94, β-His146 and α-Lys40. Methods for using these compositions, such as for treating carbon monoxide poisoning, and methods for producing these compositions, are also disclosed.

Compositions that include a globin, such as hemoglobin, in a relaxed state are described. Globin molecules in a relaxed state (R state) have a higher binding affinity for carbon monoxide and oxygen than globin molecules in a tense state (T state). Hemoglobin in a relaxed state can be, for example, hemoglobin that is substantially free of 2,3-diphosphoglycerate or hemoglobin that includes a β-Cys93 that is covalently modified to inhibit one or both salt bridges between β-Asp94, β-His146 and α-Lys40. Methods for using these compositions, such as for treating carbon monoxide poisoning, and methods for producing these compositions, are also disclosed.

This document relates to meat replica food product, and more particularly to hybrid meat substitute food products comprising plant-based protein and animal cells. The meat replica mimics animal meat, including the texture, beefy flavor, and red-to-brown color transition during cooking.