CSIRO Publishing blank image blank image blank image blank imageBooksblank image blank image blank image blank imageJournalsblank image blank image blank image blank imageAbout Usblank image blank image blank image blank imageShopping Cartblank image blank image blank image You are here: Journals > Reproduction, Fertility and Development   
Reproduction, Fertility and Development
  Vertebrate Reproductive Science & Technology
 
blank image Search
 
blank image blank image
blank image
 
  Advanced Search
   

Journal Home
About the Journal
Editorial Board
Contacts
Content
All Issues
Special Issues
Research Fronts
Sample Issue
For Authors
General Information
Instructions to Authors
Submit Article
Open Access
For Referees
Referee Guidelines
Review an Article
Annual Referee Index
For Subscribers
Subscription Prices
Customer Service
Print Publication Dates

blue arrow e-Alerts
blank image
Subscribe to our email Early Alert or RSS feeds for the latest journal papers.

red arrow Connect with us
blank image
facebook twitter youtube

red arrow Connect with SRB
blank image
facebook TwitterIcon

Affiliated Societies

RFD is the official journal of the International Embryo Transfer Society and the Society for Reproductive Biology.



Article << Previous     |     Next >>   Contents Vol 22(1)

231 SELECTION OF UBIQUITOUS AND ENDOTHELIUM-SPECIFIC PROMOTERS FOR EXPRESSING TRANSGENES IN PIG

Y. Chen A and B. Mir A

Indiana University, Indianapolis, IN, USA
   

Abstract
Export Citation
Print
  


Abstract

Several promoters of different origins have been used to drive expression of transgenes in pig with variable results. However, there has not been a systemic search to identify the most reliable, ubiquitous, and tissue-specific promoters, such as endothelium-specific promoters, to drive expression of transgenes with high predictability. In this study we tested 9 ubiquitous mammalian promoters, generated by assembling enhancers, minimal promoters, and 5′untranslated region of different origins (Table 1) and 5 endothelial-specific promoters: hFlt-1 (fms-like tyrosine kinase), hICAM-2 (intercellular adhesion molecule-2), andhEndoglin promoters from human; mouse Tie-2 promoter (Schlaeger et al. 1997 PNAS 94, 3058-3063); and pig ICAM-2 promoter (Godwin et al. 2006 Xenotransplantation 13, 514-521). These promoters drive the expression of either GFP or LacZ reporter genes and were electroporated into 9 different pig cell types: endothelium-derived 2A2 and PEDSV15 cells (Seebach et al. 2001 Xenotransplantation 8, 48-61), nasal fibroblasts, kidney epithelial cells, and lung macrophages (ATCC, CRL-2528, 2842, and 2843), pig fetal fibroblasts, liver stem cells, fat-derived stem cells, and immortalized liver endothelial cells (generated in the lab). The expression was graded as week, moderate, or strong depending on the brightness of GFP signal or intensity of LacZ staining. Out of 9 ubiquitous promoters, 7 showed either low expression in all cell types or different levels of expression in some of the cell types. However, 2 promoters, hCMV-hCMV-HTLV and SV40-hCMV-HTLV, showed strong and stable expression across all 9 cell types. Among endothelial-specific promoters, pig ICAM-2 promoter showed moderate and specific expression in all 3 endothelial cell types (strong expression with the enhancer) and no expression in non-endothelial cells. Human EC-specific promoter hFlt-1 showed moderate expression in all 3 endothelial cell types, as well as in some of the non-endothelial cells such as liver stem cells, fat-derived stem cells, and lung macrophages. The other endothelial-specific human and mouse promoters did not show any expression in both endothelial and non-endothelial cells. In summary, we identified hCMV-hCMV-HTLV and SV40-hCMV-HTLV as 2 ubiquitous promoters and pICAM2 as an endothelial-specific promoter for expression of transgenes in pig. In the future, we will test these 3 promoters for in vivo expression studies.

Reproduction, Fertility and Development 22(5305) 273–274   http://dx.doi.org/10.1071/RDv22n1Ab231
Published online: 08 December 2009




 
Top  Email this page
 
   


Legal & Privacy | Contact Us | Help

CSIRO

© CSIRO 1996-2014