vsv g envelope expressing vector pmd2 g  (Addgene inc)

Journal: Viruses

Article Title: Efficient Inhibition of HIV Using CRISPR/Cas13d Nuclease System

doi: 10.3390/v13091850

Figure Lengend Snippet: Nuclear localization of CasRx mediates optimal inhibition of HIV-1. Lenti-X TM cells were transfected with plasmid encoding CasRx-NLS-GFP, CasRx-NES-GFP, poly-gRNAs targeting HIV (polyHIV), or non-targeting control (polyNT). Twenty-four hours after transfections, the cells were infected with VSV-G pseudotyped HIV-iRFP. ( A ) Forty-eight hours after infection, expression of CasRx-GFP and HIV-iRFP were measured by flow cytometry; ( B ) %iRFP expressing cells in polyHIV vs. polyNT gRNAs transfected cells were compared in all experimental groups. The means ± s.e.m. are depicted as horizontal and vertical bars for each group, respectively. The Student’s t-test was used for statistical comparisons, and two-tailed p values are indicated.

Article Snippet: The CasRx, (pXR001; Addgene #109049), dCasRx (PXR002; Addgene #109050) plasmids, gRNA expression vectors (pXR003, pXR004; Addgene #109053 and #109054), Vesicular Stomatitis Virus glycoprotein (VSV-G) envelope expression vector (pMD2.G; Addgene #12259), lentiviral packaging plasmid (psPax2; Addgene # 12260), iRFP670 fluorescent reporter vector (piRFP670-N1; Addgene#79987) and pKLV2-U6gRNA5(BbsI)-PGKpuro2ABFP-W (Addgene #67974) were obtained from the Addgene repository (Watertown, MA, USA).

Techniques: Inhibition, Transfection, Plasmid Preparation, Infection, Expressing, Flow Cytometry, Two Tailed Test