title: A novel method for somatic transgenesis of the mouse prostate using the Sleeping Beauty transposon system.
abstract: BACKGROUND
  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.
  
  
  METHODS
  Mouse e16 urogenital sinuses (UGSs) were co-injected with plasmids expressing the Sleeping Beauty transposase and plasmids with control or activated BRAF 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.
  
  
  RESULTS
  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 BRAF resulted in ectopic BRAF expression that was detectable at both the mRNA and protein levels. Prostatic ducts over-expressing activated BRAF also had ectopic activation of the ERK1/2 mitogen activated kinases and increased epithelial cell proliferation.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Division of Pharmaceutical Sciences, School of Pharmacy and University of Wisconsin Carbone Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin.
authors.forename: Kimberly D P
authors.initials: KD
authors.lastname: Hammer
authors.forename: James D
authors.initials: JD
authors.lastname: Alsop
authors.forename: Rita A
authors.initials: RA
authors.lastname: Buresh-Stiemke
authors.forename: Katsiaryna
authors.initials: K
authors.lastname: Frantskevich
authors.forename: Rita L
authors.initials: RL
authors.lastname: Malinowski
authors.forename: Laura S
authors.initials: LS
authors.lastname: Roethe
authors.forename: Ginny L
authors.initials: GL
authors.lastname: Powers
authors.forename: Paul C
authors.initials: PC
authors.lastname: Marker
mesh_terms: D064591:Allografts; D000818:Animals; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D011467:Prostate / Q000378:metabolism*; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D019895:Transposases / Q000235:genetics* / Q000378:metabolism
chemical_list: D019895:Transposases
keywords: BRAF; ERK1; ERK2; Sleeping Beauty transposon; mouse prostate; transgenic