A novel method for somatic transgenesis of the mouse prostate using the Sleeping Beauty transposon system. Issue 7 (20th March 2014)

• Record Type: Journal Article
• Authors: Hammer, Kimberly D.P.; Alsop, James D.; Buresh‐Stiemke, Rita A.; Frantskevich, Katsiaryna; Malinowski, Rita L.; Roethe, Laura S.; Powers, Ginny L.; Marker, Paul C.
• Is Part Of: Prostate. Volume 74:Issue 7(2014)
• Page Start: 781
• Page End: 791
• Publication Date: 2014-03-20
• Subjects: Prostate -- Diseases -- Periodicals; 616
• DOI: 10.1002/pros.22797
• Languages: English
• ISSNs: 0270-4137

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <sec id="pros22797-sec-0001" sec-type="section"> <title>BACKGROUND</title> <p>In vivo ectopic gene expression is a common approach for prostate research through the use of transgenes in germline transgenic mice. For some other organs, somatic transgenesis with the Sleeping Beauty transposon system has allowed in vivo ectopic gene expression with higher throughput and lower cost than germline transgenic approaches.</p> </sec> <sec id="pros22797-sec-0002" sec-type="section"> <title>METHODS</title> <p>Mouse e16 urogenital sinuses (UGSs) were co‐injected with plasmids expressing the Sleeping Beauty transposase and plasmids with control or activated <italic>BRAF</italic> expressing transposons. Following electroporation, the transduced UGSs were grown as allografts in mouse hosts for 8 weeks, and the resulting allografts were evaluated for several endpoints.</p> </sec> <sec id="pros22797-sec-0003" sec-type="section"> <title>RESULTS</title> <p>Transposon‐transduced UGS allografts developed into prostatic tissue with normal tissue structure and cellular differentiation. Integration of transposon vectors into the genomes of transduced allografts was confirmed using linker‐mediated PCR, sequencing, and in situ PCR. Transduction of UGS allografts with transposons expressing activated <italic>BRAF</italic> resulted in ectopic <italic>BRAF</italic> expression that was detectable at both the mRNA and protein levels. Prostatic ducts over‐expressing activated <italic>BRAF</italic> also had ectopic activation of the ERK1/2 mitogen activated kinases and increased epithelial cell proliferation.</p> </sec> <sec id="pros22797-sec-0004" sec-type="section"> <title>CONCLUSIONS</title> <p>The Sleeping Beauty transposon system can be used to achieve somatic transgenesis of prostatic allografts. This new method for achieving ectopic gene expression in the prostate will complement other existing approaches such as ectopic gene expression in cell lines and in germline transgenic mice. Advantages of this new approach include preservation of stromal–epithelial interactions not possible with cell lines, and higher throughput and lower cost than traditional germline transgenic approaches. <italic>Prostate 74:781–791, 2014</italic>. © 2014 Wiley Periodicals, Inc.</p> </sec> </abstract>