...single experiment wherein Plasmid A (encoding shRNA against human Gene X in backbone Y) was transfected ...variable is the shRNA. Therefore the empty control vector would be Plasmid A sans shRNA, or backbone Y ...every experiment, an investigator needs a standard against which the results can be compared; results from...Experiment: Knockdown the expression of Gene X using shRNA expressed from a plasmid   Figure 1: Expression ... this result one could simply conclude that the shRNA didn’t work, as the expression level of Gene X in...plasmid delivered into the cells successfully? Is the shRNA target correct? Is the purified plasmid DNA viable...constitutively expresses a reporter protein (e.g., GFP or luciferase) that is either co-transfected with the test ...

...sgRNA (U6) expression, shRNA/shRNA-miR30 constitutive (H1, U6, 7SK) or inducible shRNA-miR30 (CMV-TO) expression...constitutive or inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene...alternative to the widely used luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids... issues with fluorescent pH sensors, a set of luciferase-fluorophore pH fusion reporters, pRSETb-pHlash...insensitivity to other ions In BRET, A mutant Renilla luciferase, catalyzes the oxidation of its substrate, luciferin...containing Cas9, EGFP, mCherry, iRFP, tdTomato, luciferase, LacZ, puromycin or CreERT2 can be combined with...gene (EGFP, iRFP, IFP1.4, puromycin, neomycin or luciferase) to create a final lentiviral vector containing...

...Protein A-Renilla luciferase and the “prey” proteins are tagged with Firefly luciferase. The bait and prey... to screen the NCC-1 library of approved drugs against the entire kit; a parallel analysis that successfully...expressing GFP, tetracycline response elements, and shRNAs many of which are available at Addgene. Since GMAP...immunoprecipitated from the cells using the Protein A tag. The luciferase activities can then be measured and normalized... assess interactions. Higher relative firefly luciferase activity in the Co-IP compared of a control Co-IP...sgRNA expression vectors (containing ten guides against each of the 8,158 predicted T. gondii protein-coding...