Biomedical applications of HIV-1 restriction factors

Project: genome editing of restriction factors to confer resistance to HIV-1.

The human versions of restriction factors APOBEC3G and TRIM5alpha are unable to efficiently target HIV-1, because of 1) an inability of human TRIM5alpha to physically bind its target, the capsid protein of HIV-1; and 2) the degradation of human TRIM5alpha by the specialized HIV-1 viral protein Vif. My laboratory has isolated and characterized mutations of TRIM5alpha that confer restriction activity against HIV-1. Other groups have published that a single mutation in human APOBEC3G can make it resistant to HIV-1 Vif and thus can result in significant restriction as well. These two restriction factors act at different stages of viral replication, and it is likely that co-expressing the mutated, HIV-1-restrictive versions of TRIM5alpha and APOBEC3G would very effectively cripple the virus.
The project will thus consist in editing the human genome in cell line models in order to generate cells bearing mutations at either or both of the genes. For this, we will use genome editing approaches based on TALENs and also on the CRISPR-Cas system. Resistance of the cell lines created to HIV-1 will then be characterized by infection assays (potency, specificity, etc.). We will also investigate possible HIV-1 resistance to these tailored restriction factors, as well as the mechanism of restriction itself. We are also working at improving the efficiency of genome editing protocols, especially the homologous recombination step, through various strategies. We collaborate with partners for developing these projects toward in vivo applications, and the selected student might also work at this side of the project.
Interested students should contact Dr Berthoux for more information (Berthoux@uqtr.ca).

Faculty Supervisor:

Lionel Berthoux

Student:

CINTHIA VIOLETA HERNANDEZ PUENTE

Partner:

Discipline:

Biology

Sector:

University:

Program:

Globalink