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 DNA format of this control construct permits direct transfection in knockdown experiments or use in the viral packaging process (for lentiviral-based knockdown experiments).
The MISSION Beta-2-Microglobulin (B2M) shRNA Control Vector is a lentiviral-based plasmid vector that contains a short hairpin sequence targeting the human Beta-2-Microglobulin transcript (Gene Symbol: B2M). Beta-2-microglobulin is a MHC Class I molecule present on most cell types. It is commonly used as an endogenous control due to this universal expression. The vector backbone is the same as the MISSION TRC shRNA collections (pLKO.1). The B2M clone (TRCN0000057254) was chosen from the available complete target set (NM_004048) and has produced consistent knockdown of 80% compared to normal gene expression in the human epithelial lung carcinoma cell line A549 via quantitative RT PCR analysis.
Ampicillin and puromycin antibiotic resistance genes provide selection in bacterial or mammalian cells respectively. In addition, self-inactivating replication incompetent viral particles can be produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids (Product No. SHP001). The MISSION Beta-2-Microglobulin (B2M) shRNA Control Vector is provided as 10 μg of plasmid DNA in Tris-EDTA (TE) buffer at a concentration of 500 ng/μl.
To see more application data, protocols, vector maps visit sigma.com/shrna.
Journal of virology, 83(7), 2839-2850 (2009-01-16)
Human cytomegalovirus (HCMV), a member of the beta subgroup of the family Herpesviridae, causes serious health problems worldwide. HCMV gene expression in host cells is a well-defined sequential process: immediate-early (IE) gene expression, early-gene expression, DNA replication, and late-gene expression....
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