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Hultquist et al. report a high-throughput platform for the efficient, multiplex editing
of host factors that control HIV infection in primary CD4+ T cells. Arrayed electroporation
of CRISPR/Cas9 ribonucleoproteins (RNPs) permits the rapid generation of isogenic
human cells with ablated candidate factors and identifies gene modifications that
provide viral resistance.
Here, the scientists adapted this methodology to a high-throughput platform for the efficient, arrayed editing of candidate host factors. CXCR4 or CCR5 knockout cells generated with this method are resistant to HIV infection in a tropism-dependent manner, whereas knockout of LEDGF or TNPO3 results in a tropism-independent reduction in infection. CRISPR/Cas9 RNPs can furthermore edit multiple genes simultaneously, enabling studies of interactions among multiple host and viral factors. Finally, in an arrayed screen of 45 genes associated with HIV integrase, they identified several candidate dependency/restriction factors, demonstrating the power of this approach as a discovery platform. This technology should accelerate target validation for pharmaceutical and cell-based therapies to cure HIV infection.