oligonucleotides encoding shrna sequences  (Genechem)

Journal:

Article Title: 55K isoform of CDK9 associates with Ku70 and is involved in DNA repair

doi: 10.1016/j.bbrc.2010.05.092

Figure Lengend Snippet: A. The genomic structure of the human CDK9 gene and transcription start sites of mRNAs encoding the 55K and 42K proteins are shown (24). The sequence of the wild type 55K mRNA and the sequence of the mRNA encoding the shRNA-resistant 55K protein, termed 55Kr, are shown. B. At four days post-infection, HeLa cell extracts were prepared from cultures infected in duplicate with 55K-shRNA lentivirus, parental Vector-shRNA lentivirus; a single culture was mock-infected. Expression levels of the indicated proteins were examined in immunoblots. C. Expression of the eGFP reporter protein in the cultures infected with the indicated shRNA viruses was examined by flow cytometry at two, four, and seven days post-infection (top panel). Portions of cell cultures were also stained with propidium iodide; the percentage of cells that had a sub-2N DNA content as determined by flow cytometry are shown (bottom panel).

Article Snippet: DNA oligonucleotides encoding shRNA sequences were purchased from Invitrogen, and inserted under the control of the U6 promoter in the FG12 lentiviral vector ( 12 ).

Techniques: Sequencing, shRNA, Infection, Plasmid Preparation, Expressing, Western Blot, Flow Cytometry, Staining

Journal:

Article Title: 55K isoform of CDK9 associates with Ku70 and is involved in DNA repair

doi: 10.1016/j.bbrc.2010.05.092

Figure Lengend Snippet: A. HeLa cells were infected with the 55K-shRNA or Vector-shRNA lentiviruses and at two days post-transduction were processed for indirect immunofluorescence to detect γ-H2AX to monitor DSBs and stained with DAPI to visualize DNA. B. Pools of puromycin-resistant HeLa cells over-expressing the wild type 55K CDK9 (puro55K-WT) or the shRNA-resistant CDK9 protein (Puro55Kr) were infected with the 55K-shRNA, Con-shRNA, or parental lentiviral vector At day four post-infection cell extracts were prepared and the indicated proteins evaluated in immunoblots.

Article Snippet: DNA oligonucleotides encoding shRNA sequences were purchased from Invitrogen, and inserted under the control of the U6 promoter in the FG12 lentiviral vector ( 12 ).

Techniques: Infection, shRNA, Plasmid Preparation, Transduction, Immunofluorescence, Staining, Expressing, Western Blot