harnessing a high cargo-capacity transposon for genetic applications in vertebrates利用高舰队转座子基因在脊椎动物中的应用.pdfVIP

Harnessing a High Cargo-Capacity Transposon for Genetic Applications in Vertebrates 1,2 1,3[ 1,4[ 1 1,2,5 1,2 Darius Balciunas , Kirk J. Wangensteen , Andrew Wilber , Jason Bell , Aron Geurts , Sridhar Sivasubbu , Xin Wang1,6, Perry B. Hackett1,2,4,6, David A. Largaespada1,2,5, R. Scott McIvor1,2,4,5, Stephen C. Ekker1,2,3,5,6* 1 The Arnold and Mabel Beckman Center for Transposon Research, Institute of Human Genetics, University of Minnesota, Minneapolis, Minnesota, United States of America, 2 Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, Minnesota, United States of America, 3 Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, Minnesota, United States of America, 4 Gene Therapy Program, University of Minnesota, Minneapolis, Minnesota, United States of America, 5 Cancer Center, University of Minnesota, Minneapolis, Minnesota, United States of America, 6 Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, United States of America Viruses and transposons are efficient tools for permanently delivering foreign DNA into vertebrate genomes but exhibit diminished activity when cargo exceeds 8 kilobases (kb). This size restriction limits their molecular genetic and biotechnological utility, such as numerous therapeutically relevant genes that exceed 8 kb in size. Furthermore, a greater payload capacity vector would accommodate more sophisticated cis cargo designs to modulate the expression and mutagenic risk of these molecular therapeutics. We show that the Tol2 transposon can efficiently integrate DNA sequences larger than 10 kb into human cells. We characterize