Abstract
Mouse telokin and SM22α promoters have previously been shown to direct smooth muscle cell-specific expression of transgenes in vivo in adult mice. However, the activity of these promoters is highly dependent on the integration site of the transgene. In the current study, we found that the ectopic expression of telokin promoter transgenes could be abolished by flanking the transgene with insulator elements from the H19 gene. However, the insulator elements did not increase the proportion of mouse lines that exhibited consistent, detectable levels of transgene expression. In contrast, when transgenes were targeted to the hprt locus, both telokin and SM22α promoters resulted in reproducible patterns and levels of transgene expression in all lines of mice examined. Telokin promoter transgene expression was restricted to smooth muscle tissues in adult and embryonic mice. As reported previously, SM22α transgenes were expressed at high levels specifically in arterial smooth muscle cells; however, in contrast to randomly integrated transgenes, the hprt-targeted SM22α transgenes were also expressed at high levels in smooth muscle cells in veins, bladder, and gallbladder. Using hprt-targeted transgenes, we further analyzed elements within the telokin promoter required for tissue specific activity in vivo. Analysis of these transgenes revealed that the CArG element in the telokin promoter is required for promoter activity in all tissues and that the CArG element and adjacent AT-rich region are sufficient to drive transgene expression in bladder but not intestinal smooth muscle cells.
| Original language | English (US) |
|---|---|
| Pages (from-to) | C1024-C1032 |
| Journal | American Journal of Physiology - Cell Physiology |
| Volume | 292 |
| Issue number | 3 |
| DOIs | |
| State | Published - Mar 1 2007 |
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Keywords
- CArG element
- Development
- Embryos
- Myosin light chain kinase
- Visceral smooth muscle
ASJC Scopus subject areas
- Clinical Biochemistry
- Cell Biology
- Physiology
Cite this
Hprt-targeted transgenes provide new insights into smooth muscle-restricted promoter activity. / Touw, Ketrija; Hoggatt, April M.; Simon, Gina; Herring, B. Paul.
In: American Journal of Physiology - Cell Physiology, Vol. 292, No. 3, 01.03.2007, p. C1024-C1032.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Hprt-targeted transgenes provide new insights into smooth muscle-restricted promoter activity
AU - Touw, Ketrija
AU - Hoggatt, April M.
AU - Simon, Gina
AU - Herring, B. Paul
PY - 2007/3/1
Y1 - 2007/3/1
N2 - Mouse telokin and SM22α promoters have previously been shown to direct smooth muscle cell-specific expression of transgenes in vivo in adult mice. However, the activity of these promoters is highly dependent on the integration site of the transgene. In the current study, we found that the ectopic expression of telokin promoter transgenes could be abolished by flanking the transgene with insulator elements from the H19 gene. However, the insulator elements did not increase the proportion of mouse lines that exhibited consistent, detectable levels of transgene expression. In contrast, when transgenes were targeted to the hprt locus, both telokin and SM22α promoters resulted in reproducible patterns and levels of transgene expression in all lines of mice examined. Telokin promoter transgene expression was restricted to smooth muscle tissues in adult and embryonic mice. As reported previously, SM22α transgenes were expressed at high levels specifically in arterial smooth muscle cells; however, in contrast to randomly integrated transgenes, the hprt-targeted SM22α transgenes were also expressed at high levels in smooth muscle cells in veins, bladder, and gallbladder. Using hprt-targeted transgenes, we further analyzed elements within the telokin promoter required for tissue specific activity in vivo. Analysis of these transgenes revealed that the CArG element in the telokin promoter is required for promoter activity in all tissues and that the CArG element and adjacent AT-rich region are sufficient to drive transgene expression in bladder but not intestinal smooth muscle cells.
AB - Mouse telokin and SM22α promoters have previously been shown to direct smooth muscle cell-specific expression of transgenes in vivo in adult mice. However, the activity of these promoters is highly dependent on the integration site of the transgene. In the current study, we found that the ectopic expression of telokin promoter transgenes could be abolished by flanking the transgene with insulator elements from the H19 gene. However, the insulator elements did not increase the proportion of mouse lines that exhibited consistent, detectable levels of transgene expression. In contrast, when transgenes were targeted to the hprt locus, both telokin and SM22α promoters resulted in reproducible patterns and levels of transgene expression in all lines of mice examined. Telokin promoter transgene expression was restricted to smooth muscle tissues in adult and embryonic mice. As reported previously, SM22α transgenes were expressed at high levels specifically in arterial smooth muscle cells; however, in contrast to randomly integrated transgenes, the hprt-targeted SM22α transgenes were also expressed at high levels in smooth muscle cells in veins, bladder, and gallbladder. Using hprt-targeted transgenes, we further analyzed elements within the telokin promoter required for tissue specific activity in vivo. Analysis of these transgenes revealed that the CArG element in the telokin promoter is required for promoter activity in all tissues and that the CArG element and adjacent AT-rich region are sufficient to drive transgene expression in bladder but not intestinal smooth muscle cells.
KW - CArG element
KW - Development
KW - Embryos
KW - Myosin light chain kinase
KW - Visceral smooth muscle
UR - http://www.scopus.com/inward/record.url?scp=33947302559&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33947302559&partnerID=8YFLogxK
U2 - 10.1152/ajpcell.00445.2006
DO - 10.1152/ajpcell.00445.2006
M3 - Article
C2 - 17079332
AN - SCOPUS:33947302559
VL - 292
SP - C1024-C1032
JO - American Journal of Physiology
JF - American Journal of Physiology
SN - 0363-6143
IS - 3
ER -