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Efficient mRNA delivery to resting T cells to reverse HIV latency
Resting T cells are difficult to manipulate, and are a reservoir for latent HIV. Here, the authors develop a lipid nanoparticle formulation with the ability to transfect resting primary human T cells, enabling delivery of mRNAs that result in reactivation of latent HIV. This could help development of HIV cure strategies.
Combined, these findings demonstrate that Tat-LNP X is a highly potent latency-reversing agent that can overcome HIV RNA transcription and processing blocks to elongation, splicing and completion, and induce viral protein expression, all in the absence of T cell activation. Interestingly, the high level of latency reversal by Tat-LNP X appeared insufficient to drive virus-mediated cytotoxicity ex vivo, consistent with latently infected cells overexpressing pro-survival proteins such as B-Cell Lymphoma (BCL)-2 43, 44 and BIRC-5 45. Reporter mRNA encoding mScarlet was synthesised using in vitro transcription from codon-optimised PCR template using HiScribe T7 High Yield RNA Synthesis Kit (New England Biolabs), replacing all uridine with N1-methylpseudouridine (TriLink Biotechnologies).
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