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Engineered transposon vectors for the transfer of large gene constructs

A 14kb piece of DNA, flanked by a pair of Paris elements, appears to have transposed in Drosophila virilis. We hypothesized that this kind of “sandwich” arrangements of two complete SB elements flanking a transgene will increase the ability of the vector to transpose larger pieces of DNA. Thus, we flanked an approximately 5 kb piece of DNA with two intact copies of SB in an inverted orientation. The vector was designed in a way that transposase was able to bind to its internal binding sites within each element but its ability to cleave DNA at those sites was abolished. Efficiency of transposition of the sandwich element was about 4-fold increased compared to an SB vector containing the same marker gene. Thus, the sandwich transposon vector can be useful to extend the cloning capacity of SB elements for the transfer of large genes whose stable integration into genomes has been problematic with current viral and nonviral vectors.

Reported by

Max Delbruck Center for Molecular Medicine
Robert Rossle Str. 10
13092 Berlin
Germany
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