冷泉港免费发布siRNA等前沿技术设计方案

　　

冷泉港八月精选实验方案设计手册发布最新前沿技术设计方案，首先推荐的是Salk研究所Inder Verma教授的siRNA设计手册，第二个是杜克大学华裔博士王小凡的癌症细胞渗透研究细胞外基质研究方案。

 

据纽约冷泉生物港实验室报道，使用慢病毒做载体携带小RNA沉默基因表达的siRNA（interfering RNA）技术为科学家们研究基因功能打开一扇新的窗口，科学家们可在活体或是体外研究基因功能。冷泉港实验室出版社在8月杂志上发表一对来自Salk Institute美国癌症协会教授Inder Verma发布的siRNA设计方案，Inder Verma一直致力于siRNA疗法的研究，以慢病毒为载体携带起调节作用的RNA片段，新设计的siRNA可应用于多种细胞，并对特定基因可产生持久的沉默作用。冷泉港生物实验室新推出的siRNA设计方案在冷泉港官网免费开放，详情请点击

http://www.cshprotocols.org/cgi/content/full/2008/9/pdb.prot5009。

原文摘要：Design and Cloning of an shRNA into a Lentiviral Silencing Vector: Version A。

 

Inder M. Verma简介

Salk研究所遗传学实验室分子生物学教授，美国癌症学会会员，致力于研究携带基因药物的病毒载体的研究。

 

主要的研究方向：

HIV基因疗法

病毒载体携带凝固基因治疗先天缺陷小鼠

乳腺癌两个基因家族，BRCA1和BRCA2，最近Verma教授发现两个基因在细胞分裂过程中有重要作用，BRCA1还调控基因活性。

 

相关论文：

    PNAS article by Salk scientists wins 2007 Cozzarelli Prize, February 21, 2008

    Dr. Inder Verma Named Recipient of the 2008 Vilcek Foundation Prize, February 04, 2008

    New chimeric mouse model for human liver diseases, drug testing, December 03, 2007

    Salk researchers successfully deliver protein across the blood-brain barrier, May 15, 2007

    Salk scientists named 2006 AAAS Fellows, November 29, 2006

    Mouse study reveals human X-SCID gene therapy poses substantial cancer risk, April 26, 2006

    "Fail safe" mechanism that helps keep inflammation in check, August 23, 2005

    Inflammation's Trigger Finger, July 21, 2004

    Male Sex Hormones Cooperate With Breast Cancer Gene To Suppress Tumors, Salk Scientists Find, June 09, 2003

    Gene Transfer Reduces Levels of Key Alzheimer's Disease Protein, March 25, 2003

    Gene Therapy Reverses Male Infertility In Salk-led Study, May 27, 2002

    Mice Cured Of Hemophilia By Salk Gene Therapy Protocol, March 29, 1999

 

 

冷泉港八月推荐的第二个实验方案是，体外检测细胞外基质蛋白调节渗出过程的方案。体外检测渗出过程技术为研究癌细胞穿透血管壁提供技术支持，癌细胞渗透机制是癌症转移的关键机制。该体外检测技术方案由杜克大学华裔博士王小凡（生物通译，xiao-Fan Wang）提供，这套实验方案包括两个检测法检验细胞外基质蛋白在癌细胞转移过程中的作用。相关的文章，在冷泉港实验室网站可免费获得全文，(http://www.cshprotocols.org/cgi/content/full/2008/9/pdb.prot5034).

 

王小凡简介

杜克大学药理和癌症生物学博士

 

主要研究方向

癌症和免疫缺陷疾病分子机制

TGF- ß在细胞培养和动物模型中的信号机制

细胞在DNA损伤后复制和分裂检验机制，关键研究哺乳动物细胞的几个检验蛋白重要成分ATM, ATR 和 Rad17

肿瘤发生机制和肿瘤转移机制

 

相关论文

Bao, S., Tibbetts, R. S., Brumbaugh, K. M., Fang, Y., Richardson, D., Ali, A., Chen, S., Abraham, R. T., Wang, X.-F. (2001) Requirement for ATR/ATM-mediated phosphorylation of human Rad17 is required for genotoxic stress responses. Nature, 411, 969-974

Frederick, J., Liberati, N., Waddell, D., Shi, Y., Wang, X.-F. (2004) TGF-ß mediated transcriptional repression of c-myc is dependent on direct binding of Smad3 to a novel Repressive Smad Binding Element. Mol Cell Biol., 24, 2546-2559.

Ali, A., Zhang, J., Bao, S., Liu, Y., Otterness, D., Abraham, R. T., Wang, X.-F. (2004) Protein phosphatase 5 regulates ATM kinase activity during cellular responses to genotoxic stress. Genes & Dev., 18, 249-254.

Shao, R., Bao, S., Bai, X., Blanchette, C., Anderson, R. M., Marks, J. R., Wang, X.-F. (2004) Acquired expression of a mesenchyme-specific gene periostin by epithelial cancers promotes tumor angiogenesis. Mol. Cell Biol., 24, 3992-4003.

Bao, S., Ouyang, G., Huang, Z., Bai, X., Liu, M., Ma, C., Shao, R. Anderson, R. M., Wang, X.-F. (2004) Differential expression of periostin in colon cancers promotes tumor metastasis by activating AKT/PKB to enhance endothelial and cancer cells survival. Cancer Cell, 5, 329-339.

Waddell, D., Liberati, N., Frederick, J., Wang, X.-F. (2004) Casein kinase Ie plays a functional role in the transforming growth factor ß signaling pathway. J. Biol. Chem., 279, 29236-29246.

Fang, Y., Tsao, C., Goodman, B., Furumei, R., Tirado, C., Abraham, R., Wang, X.-F. (2004) ATR functions as a gene dosage-dependent tumor suppressor on a mismatch repair deficient background. EMBO J., 23, 3164-3174.

Hjelmeland, A. B., Schilling, S., Guo, X., Quarles, D., Wang, X.-F . (2005) Loss of Smad3-mediated negative regulation of Cbfa1 activity leads to an alteration in cell fate determination. Mol. Cell Biol., 25, 9460-9468.

Zhang, J., Bao, S., Kucera, K., Dean, N., Wang, X.-F. (2005) DNA damage-induced ATR activation is mediated by protein phosphatase 5. Mol. Cell Biol., 25, 9910-9919.

Jian, H., Shen, X., Liu, I. , Semenov, M., He, X., Wang, X.-F. (2006) TGF-ß induces ß-catenin nuclear translocation in a Smad3-dependent manner in human mesenchymal stem cells. Genes & Dev., 20, 666-674.

Guo, X., Ramirez, A.,Waddell, D., Li, Z., X. Liu, Wang, X.-F. (2008) Axin and GSK3ß control Smad3 protein stability and modulate TGF-ß signaling. Genes & Dev., 22, 106-120.

Ma, C., Rong, Y., Radiloff, D., Datto, M., Centeno, B., Bao, S., Chen, A., Lin, F., Jiang, S., Yeatman, T., Wang, X.-F. (2008) Extracellular matrix protein ßig-h3/TGFBI promotes metastasis of colon cancer by enhancing cell extravasation. Genes & Dev., in press.