SANTA CRUZ BIOTECHNOLOGY, INC. RBM33 shRNA Plasmid (h): sc-89462-SH BACKGROUND STORAGE AND RESUSPENSION Proteins containing RNA recognition motifs, including various hnRNP proteins, Store lyophilized shRNA plasmid DNA at 4° C with desiccant. Stable for are implicated in the regulation of alternative splicing and protein components at least one year from the date of shipment. Once resuspended, store at of snRNPs. The RBM (RNA-binding motif) gene family encodes proteins with 4° C for short term storage or -80° C for long term storage. Avoid repeated an RNA binding motif that have been suggested to play a role in the modula- freeze thaw cycles. tion of apoptosis. RBM33 (RNA-binding motif protein 33) is a 1,170 amino Resuspend lyophilized shRNA plasmid DNA in 200 µl of the deionized water acid protein that contains one RRM (RNA recognition motif) domain. Existing provided. Resuspension of the shRNA plasmid DNA in 200 µl of deionized as two alternatively splice isoforms, the RBM33 gene is conserved in chim- water makes a 0.1 µg/µl solution in a 10 mM Tris, 1 mM EDTA buffered panzee, cow, mouse, rat, chicken and zebrafish, and maps to human chromo- solution. some 7q36.3. Chromosome 7 is about 158 million bases long, encodes over 1000 genes and makes up about 5% of the human genome. The deletion of APPLICATIONS a portion of the q arm of chromosome 7 is associated with Williams-Beuren syndrome, a condition characterized by mild mental retardation, an unusual RBM33 shRNA Plasmid (h) is recommended for the inhibition of RBM33 comfort and friendliness with strangers and an elfin appearance. Deletions of expression in human cells. portions of the q arm of chromosome 7 are also seen in a number of myeloid disorders including cases of acute myelogenous leukemia and myelodysplasia. SUPPORT REAGENTS For optimal shRNA Plasmid transfection efficiency, Santa Cruz Biotechnology’s REFERENCES shRNA Plasmid Transfection Reagent: sc-108061 (0.2 ml) and shRNA Plasmid 1. Varani, G., et al. 1998. RNA recognition by RNP proteins during RNA pro- Transfection Medium: sc-108062 (20 ml) are recommended. Control shRNAs cessing. Annu. Rev. Biophys. Biomol. Struct. 27: 407-445. are available as 20 µg lyophilized plasmid DNA. Each encodes a scrambled shRNA sequence that will not lead to the specific degradation of any known 2. Hillier, L.W., et al. 2003. The DNA sequence of human chromosome 7. cellular mRNA. Control shRNA Plasmids include: sc-108060, sc-108065 and Nature 424: 157-164. sc-108066. 3. Scherer, S.W., et al. 2003. Human chromosome 7: DNA sequence and biology. Science 300: 767-772. RT-PCR REAGENTS 4. Maris, C., et al. 2005. The RNA recognition motif, a plastic RNA-binding Semi-quantitative RT-PCR may be performed to monitor RBM33 gene expres- platform to regulate post-transcriptional gene expression. FEBS J. 272: sion knockdown using RT-PCR Primer: RBM33 (h)-PR: sc-89462-PR (20 µl). 2118-2131. Annealing temperature for the primers should be 55-60° C and the extension temperature should be 68-72° C. 5. Sutherland, L.C., et al. 2005. RNA binding motif (RBM) proteins: a novel family of apoptosis modulators? J. Cell. Biochem. 94: 5-24. RESEARCH USE 6. Eckert, M.A., et al. 2006. The neurobiology of Williams syndrome: cascading The purchase of this product conveys to the buyer the nontransferable right influences of visual system impairment? Cell. Mol. Life Sci. 63: 1867-1875. to use the purchased amount of the product and all replicates and derivatives 7. Fukuda, T., et al. 2009. hnRNP K interacts with RNA binding motif pro- for research purposes conducted by the buyer in his laboratory only (whether tein 42 and functions in the maintenance of cellular ATP level during the buyer is an academic or for-profit entity). The buyer cannot sell or other- stress conditions. Genes Cells 14: 113-128. wise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party, or otherwise use this product CHROMOSOMAL LOCATION or its components or materials made using this product or its components for Commercial Purposes. Genetic locus: RBM33 (human) mapping to 7q36.3. PROTOCOLS PRODUCT See our web site at www.scbt.com for detailed protocols and support RBM33 shRNA Plasmid (h) is a pool of 3 target-specific lentiviral vector products. plasmids each encoding 19-25 nt (plus hairpin) shRNAs designed to knock down gene expression. Each plasmid contains a puromycin resistance gene for the selection of cells stably expressing shRNA. Each vial contains 20 µg of lyophilized shRNA plasmid DNA. Suitable for up to 20 transfections. Also see RBM33 siRNA (h): sc-89462 and RBM33 shRNA (h) Lentiviral Particles: sc-89462-V as alternate gene silencing products. Santa Cruz Biotechnology, Inc. 1.800.457.3801 831.457.3800 fax 831.457.3801 Europe +00800 4573 8000 49 6221 4503 0 www.scbt.com.