Generation of Cleavage-Deficient HIV Env

The vesicular stomatitis virus G (VSV-G)-encoding plasmid was previously described [73 (link)]. Transmitted/Founder (T/F) infectious molecular clones (IMCs) of patients CH058 and CH077 were previously reported [74 (link),75 (link),76 (link),77 (link)]. To generate IMCs encoding for cleavage-deficient Env, two mutations (R508S/R511S) were introduced in the furin cleavage site (⁵⁰⁸REKR⁵¹¹) using the QuikChange II XL site-directed mutagenesis protocol (Agilent Technologies, Santa Clara, CA). The presence of the desired mutations was determined by automated DNA sequencing.

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Prévost J., Medjahed H., Vézina D., Chen H.C., Hahn B.H., Smith AB I.I.I, & Finzi A. (2021). HIV-1 Envelope Glycoproteins Proteolytic Cleavage Protects Infected Cells from ADCC Mediated by Plasma from Infected Individuals. Viruses, 13(11), 2236.

Publication 2021

QuikChange II XL site-directed mutagenesis protocol

Cleavage Clones Furin Infectious Mutations Patients Plasmid Site directed mutagenesis Vesicular stomatitis virus

Corresponding Organization : Université de Montréal

Other organizations : University of Pennsylvania

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Variable analysis

independent variables

Mutations (R508S/R511S) introduced in the furin cleavage site (⁵⁰⁸REKR⁵¹¹) of the Env gene

dependent variables

Presence of the desired mutations determined by automated DNA sequencing

control variables

The vesicular stomatitis virus G (VSV-G)-encoding plasmid
Transmitted/Founder (T/F) infectious molecular clones (IMCs) of patients CH058 and CH077

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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