Generation of SIV-resistant CD4+ t cells modified with CCR5 specific Zinc-Finger Nucleases in the rhesus macaque

In this project, we wanted to see if it was possible to engineer cells that could mimic some of what was seen in the Berlin Patient. The Berlin Patient was an HIV positive man diagnosed with AML that needed bone marrow transplants to live. His doctor decided it could be beneficial to see if there was a bone marrow donor that was possibly CCR5 negative and transplant that bone marrow into the patient. They were successful in finding a donor and after three years, the patient is found to have no detectable levels of HIV in his blood stream or other reservoirs where it is usually found in an infected individual, this is even after removing HAART treatments.

Our goal was to use a style of gene editing using Zinc-Finger Nucleases (ZFN) that could delete the CCR5 region of the gene to have cells that lacked the necessary receptor for SIV envelope fusion. We chose to use CD4+ T cells since these are the targets of HIV and SIV as well as using CD34+ cells isolated from rhesus macaque bone marrow. The long-term goal of this idea is to have engineered SIV resistant cells that could possibly be transplanted back into the monkey and see if SIV resistance was possible.