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Rlf Shrna Plasmid (H): Sc-88789-SH SANTA CRUZ BIOTECHNOLOGY, INC. Rlf shRNA Plasmid (h): sc-88789-SH BACKGROUND PRODUCT Chromosome 1 is the largest human chromosome spanning about 260 million Rlf shRNA Plasmid (h) is a pool of 3 target-specific lentiviral vector plasmids base pairs and making up 8% of the human genome. There are about 3,000 each encoding 19-25 nt (plus hairpin) shRNAs designed to knock down gene genes on chromosome 1, and considering the great number of genes there are expression. Each plasmid contains a puromycin resistance gene for the selec- also a large number of diseases associated with chromosome 1. Notably, the tion of cells stably expressing shRNA. Each vial contains 20 µg of lyophiliz- rare aging disease Hutchinson-Gilford progeria is associated with the LMNA ed shRNA plasmid DNA. Suitable for up to 20 transfections. Also see Rlf gene which encodes Lamin A. When defective, the LMNA gene product can siRNA (h): sc-88789 and Rlf shRNA (h) Lentiviral Particles: sc-88789-V as build up in the nucleus and cause characteristic nuclear blebs. The mechanism alternate gene silencing products. of rapidly enhanced aging is unclear and is a topic of continuing exploration. The MUTYH gene is located on chromosome 1 and is partially responsible for STORAGE AND RESUSPENSION familial adenomatous polyposis. Stickler syndrome, Parkinsons, Gaucher dis- Store lyophilized shRNA plasmid DNA at 4° C with desiccant. Stable for ease and Usher syndrome are also associated with chromosome 1. A break- at least one year from the date of shipment. Once resuspended, store at point has been identified in 1q which disrupts the DISC1 gene and is linked to 4° C for short term storage or -80° C for long term storage. Avoid repeated schizophrenia. Aberrations in chromosome 1 are found in a variety of cancers freeze thaw cycles. including head and neck cancer, malignant melanoma and multiple myeloma. Resuspend lyophilized shRNA plasmid DNA in 200 µl of the deionized water REFERENCES provided. Resuspension of the shRNA plasmid DNA in 200 µl of deionized water makes a 0.1 µg/µl solution in a 10 mM Tris, 1 mM EDTA buffered 1. Watson, M.L., Kingsmore, S.F., Johnston, G.I., Siegelman, M.H., Le Beau, solution. M.M., Lemons, R.S., Bora, N.S., Howard, T.A., Weissman, I.L., McEver, R.P. and Seldin, M.F. 1990. Genomic organization of the selectin family of APPLICATIONS leukocyte adhesion molecules on human and mouse chromosome 1. J. Exp. Med. 172: 263-272. Rlf shRNA Plasmid (h) is recommended for the inhibition of Rlf expression in human cells. 2. Blackwood, D.H., Fordyce, A., Walker, M.T., St Clair, D.M., Porteous, D.J. and Muir, W.J. 2001. Schizophrenia and affective disorders—cosegrega- SUPPORT REAGENTS tion with a translocation at chromosome 1q42 that directly disrupts brain- expressed genes: clinical and P300 findings in a family. Am. J. Hum. Genet. For optimal shRNA Plasmid transfection efficiency, Santa Cruz Biotechnology’s 69: 428-433. shRNA Plasmid Transfection Reagent: sc-108061 (0.2 ml) and shRNA Plasmid Transfection Medium: sc-108062 (20 ml) are recommended. Control shRNAs 3. Weise, A., Starke, H., Mrasek, K., Claussen, U. and Liehr, T. 2005. New are available as 20 µg lyophilized plasmid DNA. Each encodes a scrambled insights into the evolution of chromosome 1. Cytogenet. Genome Res. shRNA sequence that will not lead to the specific degradation of any known 108: 217-222. cellular mRNA. Control shRNA Plasmids include: sc-108060, sc-108065 and 4. Lans, H. and Hoeijmakers, J.H. 2006. Cell biology: aging nucleus gets out sc-108066. of shape. Nature 440: 32-34. RT-PCR REAGENTS 5. Gregory, S.G., Barlow, K.F., McLay, K.E., Kaul, R., Swarbreck, D., Dunham, A., Scott, C.E., Howe, K.L., Woodfine, K.C., Spencer, C.A., Jones, M.C., Semi-quantitative RT-PCR may be performed to monitor Rlf gene expression Gillson, C., Searle, S., Zhou, Y., Kokocinski, F., McDonald, L., et al. 2006. knockdown using RT-PCR Primer: Rlf (h)-PR: sc-88789-PR (20 µl). Annealing The DNA sequence and biological annotation of human chromosome 1. temperature for the primers should be 55-60° C and the extension tempera- Nature 441: 315-321. ture should be 68-72° C. 6. Hennah, W., Thomson, P., Peltonen, L. and Porteous, D. 2006. Genes and RESEARCH USE schizophrenia: beyond schizophrenia: the role of DISC-1 in major mental The purchase of this product conveys to the buyer the nontransferable right illness. Schizophr. Bull. 32: 409-416. to use the purchased amount of the product and all replicates and derivatives 7. Marzin, Y., Jamet, D., Douet-Guilbert, N., Morel, F., Le Bris, M.J., Morice, for research purposes conducted by the buyer in his laboratory only (whether P., Abgrall, J.F., Berthou, C. and De Braekeleer, M. 2006. Chromosome 1 the buyer is an academic or for-profit entity). The buyer cannot sell or other- abnormalities in multiple myeloma. Anticancer Res. 26: 953-959. wise transfer (a) this product (b) its components or (c) materials made using 8. McClintock, D., Gordon, L.B. and Djabali, K. 2006. Hutchinson-Gilford proge- this product or its components to a third party, or otherwise use this product ria mutant Lamin A primarily targets human vascular cells as detected by an or its components or materials made using this product or its components anti-Lamin A G608G antibody. Proc. Natl. Acad. Sci. USA 103: 2154-2159. for Commercial Purposes. CHROMOSOMAL LOCATION PROTOCOLS Genetic locus: RLF (human) mapping to 1p34.2. 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.
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