This control is intended for use as a positive control for knockdown of human gene targets when used with clones of the MISSION TRC human shRNA collections. The lentiviral format of this product permits direct transduction of target cells. These particles are produced from a 3rd generation transfer vector (pLKO.1) and are self-inactivating and replication incompetent for safety purposes.
The MISSION Rho GDP Dissociation Inhibitor (GDI) alpha (ARHGDIA) shRNA Control Transduction Particles contains a short hairpin sequence targeting the human Rho GDP Dissociation Inhibitor (GDI) alpha transcript (Gene Symbol: ARHGDIA). Rho GDP dissociation inhibitor (GDI) alpha (ARHGDIA) is a ubiquitously expressed protein that acts on human Rho GTPases including RhoA, Rac1 and Cdc42, by keeping these proteins locked in an inactive state. Complete understanding of ARHGDIA′s roles is still being elucidated but it is believed to be involved in various signal transduction pathways and cellular cytoskeleton functions. The MISSION Rho GDP Dissociation Inhibitor (GDI) alpha (ARHGDIA) shRNA Control Transduction Particles are produced from the sequence-verified lentiviral-based pLKO.1 plasmid clone (Product No. SHC009, Clone No. TRNC0000008004) chosen from the available complete target set (NM_004309) and has produced consistent knockdown of 90% compared to normal gene expression in the human epithelial lung carcinoma cell line A549 via quantitative RT PCR analysis and Western blot analysis (Antibody Product No. R3025).
MISSION Rho GDP Dissociation Inhibitor (GDI) alpha (ARHGDIA) shRNA Control Transduction Particles, as all other MISSION lentiviral format clones, are pseudotyped with an envelope G glycoprotein from Vesicular Stomatitis Virus (VSV-G), allowing transduction of a wide variety of mammalian cells. The lentiviral particles are provided as a frozen stock in a volume of 200 μl with a minimum titer of 1 x 106 TU/ml (via p24 titering assay). Upon transduction, the sequence integrates into the host cell genome and transduced cells can be selected using puromycin resistance.
When conducting experiments using MISSION® shRNA clones, the proper controls should be a key element of your experimental design to allow for accurate interpretation of knockdown results. The MISSION Control Transduction Particles are a critical positive control to monitor transduction efficiency.
To see more application data, protocols, vector maps visit sigma.com/shrna.