Liver-directed gene therapy using the sleeping beauty transposon system.
Sleeping Beauty (SB) is a transposon system genetically reconstructed from teleost fish that mediates chromosomal integration of DNA sequences by a cut-and-paste mechanism. SB has been shown to mediate transposition in a variety of cells and tissues, has been used for the generation of transgenic animals and has been tested as a vector for gene therapy in several animal models of human disease. Here, we describe methods that we have developed for testing SB-mediated transposition, first in cultured mammalian cells, and then in vivo using a combination of rapid, high-volume tail vein injection for DNA delivery to the liver along with in vivo bioluminescence imaging to monitor sustained luciferase gene expression in individual animals.
Belur
Lalitha R
LR
Arnold and Mabel Beckman Center for Transposon Research, Gene Therapy Program, Institute of Human Genetics, and Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA.
McIvor
R Scott
RS
Wilber
Andrew
A
D000818:Animals; D004251:DNA Transposable Elements / Q000235:genetics*; D005354:Fibrosarcoma / Q000235:genetics / Q000628:therapy*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism; D049449:Luminescence; D051379:Mice; D008810:Mice, Inbred C57BL; D014162:Transfection; D019895:Transposases / Q000235:genetics* / Q000378:metabolism
D004251:DNA Transposable Elements; D008156:Luciferases; D019895:Transposases; C494471:sleeping beauty transposase, human