Journal: Molecular Biotechnology

Article Title: Universal Real-Time PCR-Based Assay for Lentiviral Titration

doi: 10.1007/s12033-014-9815-4

Figure Lengend Snippet: Flow cytometry estimated for the lentiviral titer of virus carrying GFP cassette; Histograms showing GFP-positive cells, 72 h after transduction with 4-fold dilutions of lentiviral vector

Article Snippet: For these studies, we utilized the pLV-THM-GFP lentiviral vector which expresses the GFP gene under control of the H1 promoter (Plasmid #12247; Addgene; Cambridge [USA]).

Techniques: Flow Cytometry, Transduction, Plasmid Preparation

Journal: Molecular Biotechnology

Article Title: Universal Real-Time PCR-Based Assay for Lentiviral Titration

doi: 10.1007/s12033-014-9815-4

Figure Lengend Snippet: Estimation of lentiviral titer and the calculation schema for exemplary lentivirus

Article Snippet: For these studies, we utilized the pLV-THM-GFP lentiviral vector which expresses the GFP gene under control of the H1 promoter (Plasmid #12247; Addgene; Cambridge [USA]).

Techniques:

Journal: Acta Pharmacologica Sinica

Article Title: Construction of lentivirus-based inhibitor of hsa-microRNA-338-3p with specific secondary structure

doi: 10.1038/aps.2012.172

Figure Lengend Snippet: The biological functions of miR-338-3p on CRC-derived cell line. (A) SMO 3′-UTR site potentially targeted by miR-338-3p as predicted by TargetScan. (B) Western-blot analysis showing SMO protein expression in SW-620 cells. β-Actin was used as house-keeping gene to normalize SMO protein expression. Note: lane 1, Negative control; lane 2, SW-620 cells transfected with pLV-THM-control; lanes 3, SW-620 cells transfected with pLV-THM-miR-338-3p-inhibitor. The results are representative of three independent experiments. (C–E) Transwell assay showing migration ability of the blank control SW-620 cells (C), SW-620 cells transfected with pLV-THM-control (D) and SW-620 cells transfected with pLV-THM-miR-338-3p-inhibitor (E). Migrated cells found on the bottom side of the membrane were fixed and stained. Numbers of migrated cells on the membrane bottom were counted and shown at the top of each panel. Down-expression of miR-338-3p increased the expression of SMO protein in SW-620 cells, which showed obviously enhanced invasive ability in transwell assay. n=3. Mean±SD. bP<0.01 versus control group.

Article Snippet: Transfer vector construction The third generation of self-inactivating, lentivirus plasmid, pLV-THM (HIV-1-based vector, Addgene, Cambridge, MA, USA), which contains a CMV-driven enhanced green fluorescence protein (eGFP) reporter and an H 1 promoter upstream of the restriction sites ( Mlu I and Cla I) to allow for the introduction of a short-hairpin RNA (shRNA) expression cassette, was used as the transfer plasmid and was linearized by digesting the vector with the restriction enzymes.

Techniques: Derivative Assay, Western Blot, Expressing, Negative Control, Transfection, Transwell Assay, Migration, Staining