RNA as a source of transposase for Sleeping Beauty-mediated gene insertion and expression in somatic cells and tissues.
Sleeping Beauty (SB) is a DNA transposon capable of mediating gene insertion and long-term expression in vertebrate cells when co-delivered with a source of transposase. In all previous reports of SB-mediated gene insertion in somatic cells, the transposase component has been provided by expression of a co-delivered DNA molecule that has the potential for integration into the host cell genome. Integration and continued expression of a gene encoding SB transposase could be problematic if it led to transposon re-mobilization and reintegration. We addressed this potential problem by supplying the transposase-encoding molecule in the form of mRNA. We show that transposase-encoding mRNA can effectively mediate transposition in vitro in HT1080 cells and in vivo in mouse liver following co-delivery with a recoverable transposon or with a luciferase transposon. We conclude that in vitro-transcribed mRNA can be used as an effective source of transposase for SB-mediated transposition in mammalian cells and tissues.
Wilber
Andrew
A
The Arnold and Mabel Beckman Center for Transposon Research, University of Minnesota, Minneapolis, MN 55455, USA.
Frandsen
Joel L
JL
Geurts
Jennifer L
JL
Largaespada
David A
DA
Hackett
Perry B
PB
McIvor
R Scott
RS
D000818:Animals; D045744:Cell Line, Tumor; D004251:DNA Transposable Elements; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D006801:Humans; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009928:Organ Specificity / Q000235:genetics; D012333:RNA, Messenger / Q000008:administration & dosage / Q000235:genetics*; D011995:Recombination, Genetic; D019895:Transposases / Q000235:genetics*
D004251:DNA Transposable Elements; D012333:RNA, Messenger; D019895:Transposases; C494471:sleeping beauty transposase, human