HIV-1 LTR Luciferase Reporter Assay

All desalted oligonucleotides and aptamers were purchased from Sigma-Aldrich, Milan, Italy (Supplementary Table S1). The HIV-1 LTR region was inserted into the promoterless luciferase reporter vector pGL4.10-Luc2 (Promega Italia, Milan, Italy) to form the pGL4.10-LTR-Luc2 vector, as previously reported (30 (link)). The Renilla plasmid (p4.74, Promega Italia, Milan, Italy) was used as an internal control. The human enhanced green fluorescent protein-nucleolin plasmid (GFP-Nucleolin) was purchased from Addgene (Addgene, Cambridge, MA, USA). The pEGFP empty vector was used as control (Clontech, Takara Bio, Otsu, Japan).

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Tosoni E., Frasson I., Scalabrin M., Perrone R., Butovskaya E., Nadai M., Palù G., Fabris D, & Richter S.N. (2015). Nucleolin stabilizes G-quadruplex structures folded by the LTR promoter and silences HIV-1 viral transcription. Nucleic Acids Research, 43(18), 8884-8897.

Publication 2015

desalted oligonucleotides pGL4.10-Luc2 Renilla plasmid GFP-Nucleolin

Enhanced green fluorescent protein Hiv 1 ltr Human Luciferase Nucleolin Oligonucleotides Pgl4 Plasmid Promega Renilla Vector

Corresponding Organization :

Other organizations : University of Padua, University at Albany, State University of New York

Top 5 similar protocols

Variable analysis

independent variables

HIV-1 LTR region inserted into the promoterless luciferase reporter vector pGL4.10-Luc2
Human enhanced green fluorescent protein-nucleolin plasmid (GFP-Nucleolin)
PEGFP empty vector

dependent variables

Luciferase reporter activity

control variables

Renilla plasmid (p4.74) used as an internal control

positive controls

None specified

negative controls

PEGFP empty vector

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