Development of Gene Editing Strategies for CCR5 Gene in Endothelial Cells

Author:

Number of pages: 22-30
Year-Number: 2021-2

Abstract

The CRISPR system has been adapted as a promising gene editing technology that provides robust and efficient DNA alterations. It was demonstrated that it has a high adaptability and great potential to cure acquired immune deficiency syndrome (AIDS) through gene disruptions. Endothelial cells are an essential component of the vascular system, and CRISPR has emerged as a promising candidate for AIDS therapies by removing the human immunodeficiency virus (HIV) reservoir in endothelial cells. CCR5, an essential receptor for HIV infection, is expressed by endothelial cells. We developed CCR5 gRNAs and tested their effectiveness in gene targeting. We established a basic and successful transfection system, as well as on-target CCR5 mutagenesis using the endonuclease assay. These findings imply that endothelial diseases could be targeted using the CCR5 gRNA/CRISPR system, which would be a novel approach. Endothelial diseases caused by genetic mutations or HIV infection could be treated in endothelial cells using the CRISPR/Cas9-based gene therapy platform by targeting CCR5.

Keywords

Abstract

The CRISPR system has been adapted as a promising gene editing technology that provides robust and efficient DNA alterations. It was demonstrated that it has a high adaptability and great potential to cure acquired immune deficiency syndrome (AIDS) through gene disruptions. Endothelial cells are an essential component of the vascular system, and CRISPR has emerged as a promising candidate for AIDS therapies by removing the human immunodeficiency virus (HIV) reservoir in endothelial cells. CCR5, an essential receptor for HIV infection, is expressed by endothelial cells. We developed CCR5 gRNAs and tested their effectiveness in gene targeting. We established a basic and successful transfection system, as well as on-target CCR5 mutagenesis using the endonuclease assay. These findings imply that endothelial diseases could be targeted using the CCR5 gRNA/CRISPR system, which would be a novel approach. Endothelial diseases caused by genetic mutations or HIV infection could be treated in endothelial cells using the CRISPR/Cas9-based gene therapy platform by targeting CCR5.

Keywords