NYU Langone Medical Center

Sun Lab Reagents - UPII Promoter

  1. General information (2010)

    We have isolated and characterized a 3.6 kb 5’-upstream sequence of mouse uroplakin II gene, and we showed that this sequence can serve as a bladder urothelium-specific promoter in transgenic mouse studies (Lin et al., 1995). This sequence has been used to drive the mouse urothelial expression of GFP (Kerr et al., 1998), human growth factor (Kerr et al., 1998), SV40-large T antigen (Zhang et al., 1999; Cheng et al., 2003), Ha-Ras (Zhang et al., 2001; Mo et al., 2007), EGF receptor (Cheng et al., 2002). It has also been used to achieve the urothelium-specific ablation (conventional as well as inducible) of various genes including Rb and p53 (Mo et al., 2005; He et al., 2009). Such studies have enabled Dr. Xue-Ru Wu to dissect the roles of various oncogenes and tumor-suppressor genes in bladder tumorigenesis (Mo et al., 2007). This web page and associated links provide the detailed sequences and restriction maps of this mouse UPII promoter (updated from the original paper by (Lin et al., 1995)). Blasting this sequence against the mouse genome (NCBI) showed 99% identity. While the extremely minor differences may not affect the utility of this promoter in a given transgenic experiments, you should keep such differences in mind if you see an unexpected restriction enzyme pattern. To be prudent, re-sequencing a promoter before its use is highly recommended.

    See the appended pdf file for the nucleotide sequence, restriction enzymes and maps, and genomic sequence comparisons. This promoter is in a pBluescript SK(+) vector, which contains the Amp-resistant gene. To amplify, transform this into JM109 or DH5 alpha and select on Amp plates and broth. We recommend the use of KpnI as a 5’ restriction-enzyme to cut out the promoter. The XhoI and ApaI enzymes do not cut due to a disruption during the subcloning process. Use the map to figure out what enzyme you prefer for the 3' cut. Visit Stratagen.com for additional information about the vector sequence and restriction map. If you like to use this promoter in your research, have questions, or have generated useful data/publications from the use of this promoter, please let us know: Dr. Tung-Tien Sun at sunt01@nyumc.org or Dr. Xue-Ru Wu at Xue-Ru.Wu@nyumc.org\

  2. An Update (May 2017)

    In a recent 2014 PLOS One paper, Pena and colleagues (Ayala de la Pena et al., 2014) described the use of a mouse uroplakin-II (UPK II) promoter that they cloned to drive the urothelium-specific expression of K-ras oncogene. In both Materials and Methods and Discussion of their paper, the authors made reference to the original mouse uroplakin II promoter that our group cloned and characterized in 1995 (Lin et al., 1995) as being “miscloned”, thus affecting its “activity” and “specificity”. Similar statements were made by these authors elsewhere (Ayala de la Pena et al., 2011; Kanasaki et al., 2013). We wish to point out that these statements are inaccurate. Although the UPII promoter sequence we originally deposited in GenBank contained a small error in that a SacI fragment was inadvertently reversed, once corrected the complete sequence of our UPII promoter matched perfectly with the known mouse genome sequence (accession number U14421; see diagram below). Therefore, our promoter was not “miscloned” and there is nothing wrong with its “activity” and “specificity”, as is evidenced by the fact that it has been used successfully to drive the urothelial expression of many foreign genes including SV40T (Zhang et al., 1999; Cheng et al., 2003; Zhou et al., 2012), HRAS (Zhang et al., 2001; Mo et al., 2007; He et al., 2015), p53 mutant (Gao et al., 2004), cyclin D1 (Garcia-Espana et al., 2005), EGFR (Cheng et al., 2002) and survivin (Salz et al., 2005), leading to urothelial changes mimicking different steps of tumorigenesis and progression that closely resemble what happen in human. This UPII promoter has also been used successfully to drive the urothelium-specific expression of other foreign genes including NGF (Schnegelsberg et al., 2009), MCP-1 (Xu et al., 2016), OVA (Liu et al., 2007), enabling the development of several urinary tract disease models. This promoter has also allowed the urothelium-specific knockout, both constitutive and controllable, of genes including p53, RB1 and p53/RB1 (Mo et al., 2005; He et al., 2009; He et al., 2015) and PTEN (Ahmad et al., 2011).

    Feng He, Department of Urology; Xue-Ru Wu, Departments of Urology and Pathology (Xue-Ru.Wu@nyumc.org); and Tung-Tien Sun, Departments of Cell Biology, Dermatology, Biochemistry & Molecular Pharmacology, and Urology (Sunt01@nyumc.org), New York University School of Medicine, 550 First Avenue, New York, NY 10016

  3. References
  • Ahmad, I., Morton, J.P., Singh, L.B., Radulescu, S.M., Ridgway, R.A., Patel, S., Woodgett, J., Winton, D.J., Taketo, M.M., Wu, X.R., Leung, H.Y., and Sansom, O.J. (2011). beta-Catenin activation synergizes with PTEN loss to cause bladder cancer formation. Oncogene 30, 178-189 [PMC3044461].
  • Ayala de la Pena, F., Kanasaki, K., Kanasaki, M., Tangirala, N., Maeda, G., and Kalluri, R. (2011). Loss of p53 and acquisition of angiogenic microRNA profile are insufficient to facilitate progression of bladder urothelial carcinoma in situ to invasive carcinoma. J Biol Chem 286, 20778-20787 [PMC3121487].
  • Ayala de la Pena, F., Kanasaki, K., Kanasaki, M., Vong, S., Rovira, C., and Kalluri, R. (2014). Specific activation of K-RasG12D allele in the bladder urothelium results in lung alveolar and vascular defects. PLoS One 9, e95888 [PMC3997426].
  • Cheng, J., Huang, H., Pak, J., Shapiro, E., Sun, T.T., Cordon-Cardo, C., Waldman, F.M., and Wu, X.R. (2003). Allelic loss of p53 gene is associated with genesis and maintenance, but not invasion, of mouse carcinoma in situ of the bladder. Cancer Res 63, 179-185.
  • Cheng, J., Huang, H., Zhang, Z.T., Shapiro, E., Pellicer, A., Sun, T.T., and Wu, X.R. (2002). Overexpression of epidermal growth factor receptor in urothelium elicits urothelial hyperplasia and promotes bladder tumor growth. Cancer Res 62, 4157-4163.
  • Gao, J., Huang, H.Y., Pak, J., Cheng, J., Zhang, Z.T., Shapiro, E., Pellicer, A., Sun, T.T., and Wu, X.R. (2004). p53 deficiency provokes urothelial proliferation and synergizes with activated Ha-ras in promoting urothelial tumorigenesis. Oncogene 23, 687-696.
  • Garcia-Espana, A., Salazar, E., Sun, T.T., Wu, X.R., and Pellicer, A. (2005). Differential expression of cell cycle regulators in phenotypic variants of transgenically induced bladder tumors: implications for tumor behavior. Cancer Res 65, 1150-1157.
  • He, F., Melamed, J., Tang, M.S., Huang, C., and Wu, X.R. (2015). Oncogenic HRAS Activates Epithelial-to-Mesenchymal Transition and Confers Stemness to p53-Deficient Urothelial Cells to Drive Muscle Invasion of Basal Subtype Carcinomas. Cancer Res 75, 2017-2028 [PMC4433590].
  • He, F., Mo, L., Zheng, X.Y., Hu, C., Lepor, H., Lee, E.Y., Sun, T.T., and Wu, X.R. (2009). Deficiency of pRb family proteins and p53 in invasive urothelial tumorigenesis. Cancer Res 69, 9413-9421.
  • Kanasaki, K., Yu, W., von Bodungen, M., Larigakis, J.D., Kanasaki, M., Ayala de la Pena, F., Kalluri, R., and Hill, W.G. (2013). Loss of beta1-integrin from urothelium results in overactive bladder and incontinence in mice: a mechanosensory rather than structural phenotype. FASEB J 27, 1950-1961 [PMC3633821].
  • Kerr, D.E., Liang, F., Bondioli, K.R., Zhao, H., Kreibich, G., Wall, R.J., and Sun, T.-T. (1998). The bladder as a bioreactor: urothelium production and secretion of growth hormone into urine. Nature Biotechnology 16, 75-79.
  • Lin, J.H., Zhao, H., and Sun, T.-T. (1995). A tissue-specific promoter that can drive a foreign gene to express in the suprabasal urothelial cells of transgenic mice. Proceedings of the National Academy of Sciences of the United States of America 92, 679-683.
  • Liu, W., Evanoff, D.P., Chen, X., and Luo, Y. (2007). Urinary bladder epithelium antigen induces CD8+ T cell tolerance, activation, and autoimmune response. J Immunol 178, 539-546 [PMC4596412].
  • Mo, L., Cheng, J., Lee, E.Y., Sun, T.T., and Wu, X.R. (2005). Gene deletion in urothelium by specific expression of Cre recombinase. Am J Physiol Renal Physiol 289, F562-568.
  • Mo, L., Zheng, X., Huang, H.Y., Shapiro, E., Lepor, H., Cordon-Cardo, C., Sun, T.T., and Wu, X.R. (2007). Hyperactivation of Ha-ras oncogene, but not Ink4a/Arf deficiency, triggers bladder tumorigenesis. J Clin Invest 117, 314-325.
  • Salz, W., Eisenberg, D., Plescia, J., Garlick, D.S., Weiss, R.M., Wu, X.R., Sun, T.T., and Altieri, D.C. (2005). A survivin gene signature predicts aggressive tumor behavior. Cancer Res 65, 3531-3534.
  • Schnegelsberg, B., Sun, T.T., Cain, G., Bhattacharya, A., Nunn, P.A., Ford, A.P., Vizzard, M.A., and Cockayne, D.A. (2009). Overexpression of NGF in mouse urothelium leads to neuronal hyperinnervation, pelvic sensitivity and changes in urinary bladder function. Am J Physiol Regul Integr Comp Physiol.
  • Xu, S., Wang, X., Wang, Y., Lutgendorf, S., Bradley, C., Schrepf, A., Kreder, K., O'Donnell, M., and Luo, Y. (2016). Transgenic Mice Expressing MCP-1 by the Urothelium Demonstrate Bladder Hypersensitivity, Pelvic Pain and Voiding Dysfunction: A Multidisciplinary Approach to the Study of Chronic Pelvic Pain Research Network Animal Model Study. PLoS One 11, e0163829 [PMC5042429].
  • Zhang, Z.T., Pak, J., Huang, H.Y., Shapiro, E., Sun, T.T., Pellicer, A., and Wu, X.R. (2001). Role of Ha-ras activation in superficial papillary pathway of urothelial tumor formation. Oncogene 20, 1973-1980.
  • Zhang, Z.T., Pak, J., Shapiro, E., Sun, T.-T., and Wu, X.R. (1999). Urothelium-specific expression of an oncogene in transgenic mice induced the formation of carcinoma in situ and invasive transitional cell carcinoma. Cancer Research 59, 3512-3517.
  • Zhou, H., Huang, H.Y., Shapiro, E., Lepor, H., Huang, W.C., Mohammadi, M., Mohr, I., Tang, M.S., Huang, C., and Wu, X.R. (2012). Urothelial tumor initiation requires deregulation of multiple signaling pathways: implications in target-based therapies. Carcinogenesis 33, 770-780 [PMC3384072].