| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | scFvs from the previously reported human anti-HIV bNAbs PGT145, PGT128, 10E8, and VRC07-52351–53 were gene-synthesized by GenScript (Figure S3). Synthesized scFvs included additional sequences for cloning into BamHI and NheIsites ofpRRL-MND-CD19CAR-BFP whilemaintaining thereadingframe with upstream human CD8 signal peptide and downstream CAR domains (including CD8a hinge and transmembrane domains and 4-1BB and CD3z intracellular signaling domains64). Expression of viral transgenes (BFP+) and surface expression of the CD19 and VRC07-523-CARs, detected by staining cells with biotinylated Protein L (GenScript) and phycoerythrin (PE)-streptavidin (BDBiosciences),wasconfirmedby flowcytometryonanLSRII(BD Biosciences) 5–10 days after LV transduction or gene editing. | Get A Quote |
The treatment or cure of HIV infection by cell and gene therapy has been a goal for decades. Recent advances in both gene editing and chimeric antigen receptor (CAR) technology have created new therapeutic possibilities for a variety of diseases. Broadly neutralizing monoclonal antibodies (bNAbs) with specificity for the HIV envelope glycoprotein provide a promising means of targeting HIV-infected cells. Here we show that primary human T cells engineered to express anti-HIV CARs based on bNAbs (HIVCAR) show specific activation and killing of HIV-infected versus uninfected cells in the absence of HIV replication. We also show that homology-directed recombination of the HIVCAR gene expression cassette into the C... More
