Inhibition of HIV‐1 transcription in macrophages generated from induced pluripotent stem cells with CDK2‐knockdown

HIV‐1 transcription is activated by HIV‐1 Tat protein, which recruits CDK9/cyclin T1 and other host transcriptional co‐activators to the HIV‐1 promoter. We previously showed that CDK2 is also involved in regulation of HIV‐1 transcription and that a transient CDK2 knock‐down by siRNA or CDK2 inhibition with iron chelators inhibited HIV‐1 transcription. The objective of the present study was to develop, as a proof of principle, induced pluripotent stem cells (iPSC)‐derived macrophages with CDK2 knock‐down resistant to HIV‐1. We created stable 293T cells that express CDK2‐targeted shRNA and in which HIV‐1 transcription was repressed. The CDK2 knock‐down cells were reprogrammed into iPSC using simplified procedure that yielded large amount of iPSC. The iPSC were differentiated into macrophages and analyzed for HIV‐1 transcription using VSVG‐pseudotyped HIV‐1 virus. HIV‐1 transcription was inefficient in CDK2 knock‐down iPSC‐derived macrophages comparing to the macrophages differentiated from the control iPSC. Our results indicate that CDK2 can be targeted for the inhibition of HIV‐1 transcription and provide an experimental basis for a wider study to explore the role of different host cell genes in HIV‐1 transcription regulation. Thus study was supported by NIH grants 1SC1GM082325, 2G12RR003048 and 1SC1GM081192.