Presenter: Daniel Magaoay
Authors: Daniel Magaoay, Bradford Berges
Faculty Advisor: Bradford Berges
Institution: Brigham Young University
Human immunodeficiency virus (HIV) is a human pathogen that causes severe disease by infecting human T cells. High mutation rates, recombination, and short generation times underscore how HIV infections overwhelm the immune system and result in acquired immune deficiency syndrome (AIDS) [1]. The current treatment model is anti-retroviral therapy (ART), a combination of drugs that inhibit different components of the viral replication cycle. Although successful in reducing the viral load, ART is expensive, toxic to the host, and unable to eradicate the virus [2,3]. Currently, the use of CRISPR-Cas9 to target the HIV genome is being explored as an alternative treatment because it has the potential to reduce viral loads to negligible levels and prevent the development of AIDS. CRISPR-Cas9 is a two-part system that consists of a guide RNA (gRNA) that directs the Cas9 enzyme to induce double-strand breaks at a specific DNA sequence [2]. Many studies design the gRNA to target viral DNA genes to interfere with the viral replication cycle [2]. When the gRNA targets a poorly conserved region of viral DNA, the virus is able to rapidly escape CRISPR attack and remain virulent. While studies examining the effectiveness of CRISPR-Cas9 treatment are not new, many are inconclusive because the HIV gene sequence targeted by CRISPR-Cas9 mutates, allowing HIV to escape inactivation [2]. Further, few experiments have progressed beyond in vitro testing to assess the viability of CRISPR-Cas9 treatment in model organisms. While conducting trials on humanized mice is the ultimate goal of our project, our current experiments focus on the cell culture work needed to generate lentiviral vectors capable of delivering a dual-targeting CRISPR system to tissue culture cells prior to HIV challenge in vitro, to confirm that our system is not liable to viral resistance before progressing to humanized mice experiments.