Advances in Human Immune Cell Engineering with CRISPR-Cas9 

A team led by IGI Affiliate Alex Marson (UCSF) and IGI Executive Director Jennifer Doudna (UC Berkeley) has devised a new strategy to precisely modify human immune-system “T cells” using CRISPR-Cas9 technology. In their new paper, Generation of knock-in primary human T cells using Cas9 ribonucleoproteins, published in PNAS today, the team reports that they have established a programmable tool to replace specific nucleotide sequences in the genome of mature immune cells.

Using a novel approach, the scientists were able to disable a protein on the T-cell surface – CXCR4 – which can be exploited by HIV when the AIDS virus infects T cells.  By introducing the Cas9 protein pre-complexed with a guide RNA into human T cells (using transient electroporation), these active protein/RNA complexes allowed the team to delete the HIV co-receptor (CXCR4).  The protein/RNA complexes also enabled the first successful Cas9-mediated correction of specific genome sequences in human T cells.

The use of CRISPR-Cas9 technology to facilitate genome engineering in human T cells has been a major challenge for the field.   T cell genome engineering holds great promise for cell-based therapies for cancer, HIV, primary immune deficiencies, and autoimmune diseases – and this achievement provides a versatile new research tool to understand T cell function, as well as a path toward CRISPR-Cas9-based therapies for many other serious human health issues.

Press Release
In CRISPR Advance, Scientists Successfully Edit Human T Cells – Research Has Implications for Autoimmune Diseases, AIDS, and Cancer
AAAS/EurekAlert | UCSF Press Release | July 27, 2015