Site-directed integration of transgenes: transposons revisited using DNA-binding-domain technologies.
In the last 20 years, tools derived from DNA transposons have made major contributions to genetic studies from gene delivery to gene discovery. Various complementary and fairly ubiquitous DNA vehicles have been developed. Although many transposons are efficient DNA vehicles, they appear to have limited ability to target specific sequences, since all that is required at the integration locus is the presence of a short 2- to 4-bp sequence. Consequently, insertions mediated by transposon-based vectors occur somewhat randomly. In the past 5 years, strategies have emerged to enhance the site-specificity of transposon-based vectors, and to avoid random integrations. The first proposes that new target site specificity could be grafted onto a transposase by adding a new DNA-binding domain. Alternative strategies consist of indirectly targeting either the transposase or the transposon to a chosen genomic locus. The most important information available about each strategy are presented, and limitations and future prospects are discussed.
Demattei
Marie-Véronique
MV
Université François Rabelais de Tours, GICC, 37200 Tours, France.
Thomas
Xavier
X
Carnus
Elodie
E
Augé-Gouillou
Corinne
C
Renault
Sylvaine
S
D000818:Animals; D004247:DNA / Q000235:genetics*; D004251:DNA Transposable Elements; D004331:Drosophila melanogaster; D005821:Genetic Techniques; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D018626:Retroelements; D012441:Saccharomyces cerevisiae; D019076:Transgenes / Q000235:genetics*; D019895:Transposases / Q000235:genetics*
D004251:DNA Transposable Elements; D018626:Retroelements; D004247:DNA; D019895:Transposases