SANTA CRUZ BIOTECHNOLOGY, INC. TMEM50A shRNA (m) Lentiviral Particles: sc-154474-V BACKGROUND PRODUCT TMEM50A (transmembrane protein 50A), also known as small membrane pro- TMEM50A shRNA (m) Lentiviral Particles is a pool of concentrated, trans- tein 1, is a 157 amino acid protein encoded by a gene mapping to human duction-ready viral particles containing 3 target-specific constructs that chromosome 1. Chromosome 1 is the largest human chromosome spanning encode 19-25 nt (plus hairpin) shRNA designed to knock down gene expres- about 260 million base pairs and making up 8% of the human genome. There sion. Each vial contains 200 µl frozen stock containing 1.0 x 106 infectious are about 3,000 genes on chromosome 1, and considering the great number units of virus (IFU) in Dulbecco’s Modified Eagle’s Medium with 25 mM of genes there are also a large number of diseases associated with chromo- HEPES pH 7.3. Suitable for 10-20 transductions. Also see TMEM50A some 1. Notably, the rare aging disease Hutchinson-Gilford progeria is asso- siRNA (m): sc-154474 and TMEM50A shRNA Plasmid (m): sc-154474-SH as ciated with the LMNA gene which encodes Lamin A. When defective, the alternate gene silencing products. LMNA gene product can build up in the nucleus and cause characteristic nuclear blebs. The mechanism of rapidly enhanced aging is unclear and is APPLICATIONS a topic of continuing exploration. The MUTYH gene is located on chromo- TMEM50A shRNA (m) Lentiviral Particles is recommended for the inhibition some 1 and is partially responsible for familial adenomatous polyposis. of TMEM50A expression in mouse cells. Stickler syndrome, Parkinsons, Gaucher disease and Usher syndrome are also associated with chromosome 1. A breakpoint has been identified in 1q SUPPORT REAGENTS which disrupts the DISC1 gene and is linked to schizophrenia. Aberrations in chromosome 1 are found in a variety of cancers including head and neck Control shRNA Lentiviral Particles: sc-108080. Available as 200 µl frozen 6 cancer, malignant melanoma and multiple myeloma. viral stock containing 1.0 x 10 infectious units of virus (IFU); contains an shRNA construct encoding a scrambled sequence that will not lead to the REFERENCES specific degradation of any known cellular mRNA. 1. Watson, M.L., et al. 1990. Genomic organization of the selectin family of RT-PCR REAGENTS leukocyte adhesion molecules on human and mouse chromosome 1. J. Exp. Med. 172: 263-272. Semi-quantitative RT-PCR may be performed to monitor TMEM50A gene expression knockdown using RT-PCR Primer: TMEM50A (m)-PR: sc-154474-PR 2. Blackwood, D.H., et al. 2001. Schizophrenia and affective disorders— (20 µl). Annealing temperature for the primers should be 55-60° C and the cosegregation with a translocation at chromosome 1q42 that directly dis- extension temperature should be 68-72° C. rupts brain-expressed genes: clinical and P300 findings in a family. Am. J. Hum. Genet. 69: 428-433. BIOSAFETY 3. Weise, A., et al. 2005. New insights into the evolution of chromosome 1. Lentiviral particles can be employed in standard Biosafety Level 2 tissue Cytogenet. Genome Res. 108: 217-222. culture facilities (and should be treated with the same level of caution as 4. Lans, H. and Hoeijmakers, J.H. 2006. Cell biology: aging nucleus gets out with any other potentially infectious reagent). Lentiviral particles are repli- of shape. Nature 440: 32-34. cation-incompetent and are designed to self-inactivate after transduction and integration of shRNA constructs into genomic DNA of target cells. 5. Hennah, W., et al. 2006. Genes and schizophrenia: beyond schizophrenia: the role of DISC1 in major mental illness. Schizophr. Bull. 32: 409-416. STORAGE 6. Gregory, S.G., et al. 2006. The DNA sequence and biological annotation of Store lentiviral particles at -80º C. Stable for at least one year from the human chromosome 1. Nature 441: 315-321. date of shipment. Once thawed, particles can be stored at 4º C for up to 7. Marzin, Y., et al. 2006. Chromosome 1 abnormalities in multiple myeloma. one week. Avoid repeated freeze thaw cycles. Anticancer Res. 26: 953-959. RESEARCH USE 8. McClintock, D., et al. 2006. Hutchinson-Gilford progeria mutant Lamin A The purchase of this product conveys to the buyer the nontransferable right primarily targets human vascular cells as detected by an anti-Lamin A to use the purchased amount of the product and all replicates and derivatives G608G antibody. Proc. Natl. Acad. Sci. USA 103: 2154-2159. for research purposes conducted by the buyer in his laboratory only (whether 9. Scaffidi, P. and Misteli, T. 2006. Lamin A-dependent nuclear defects in the buyer is an academic or for-profit entity). The buyer cannot sell or other- human aging. Science 312: 1059-1063. 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 Genetic locus: Tmem50a (mouse) mapping to 4 D3. for Commercial Purposes. PROTOCOLS See our web site at www.scbt.com for detailed protocols and support 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.