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. 2021 Jul 22;12:663793. doi: 10.3389/fneur.2021.663793

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Copyright © 2021 Lin, Elbezanti, Schirling, Ahmed, Van Duyne, Cocklin and Klase.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Deletion of the RUNX1 binding site from the HIV-1 LTR enhances replication kinetics relative to WT. (A) J-LTR-GFP cells were transfected with pNL4-3 WT, ΔNef, ΔRUNX molecular clones and replication was monitored by measuring the percentage of GFP positive cells in the culture every 2-3 days post-transfection. (B) Jurkat cells were infected with equal nanogram quantities of viral stocks for ΔNef or ΔRUNX and replication was followed over time by p24 ELISA. (C) A competition assay was performed where J-LTR-GFP were infected with equal infectious units of the two viruses and supernatant from the peak of infection was used to infect a new round of cells. Extracellular RNA isolated from the peak of each round of infection was amplified using primers against the LTR and sequenced to be able to determine the relative abundance of each virus in each round of infection.