SITE-SPECIFIC RECOMBINATION MEDIATED BY AN ADENOVIRUS VECTOR EXPRESSING THE CRE RECOMBINASE PROTEIN - A MOLECULAR SWITCH FOR CONTROL OF GENE-EXPRESSION
M. Anton et Fl. Graham, SITE-SPECIFIC RECOMBINATION MEDIATED BY AN ADENOVIRUS VECTOR EXPRESSING THE CRE RECOMBINASE PROTEIN - A MOLECULAR SWITCH FOR CONTROL OF GENE-EXPRESSION, Journal of virology, 69(8), 1995, pp. 4600-4606
have constructed replication-defective human adenovirus (Ad) type 5 ve
ctors containing the gene for the Cre recombinase from bacteriophage P
1 under control of the human cytomegalovirus immediate-early promoter
(AdCre). Expression of the protein was detected in replication-permiss
ive (293) and in nonpermissive (MRCS) cell lines, and its biochemical
activity was demonstrated in a cell-free recombination assay using a p
lasmid containing two loxP sites. To study Cre-mediated recombination
in an intracellular system, we constructed an Ad vector (AdMA19) conta
ining the luciferase cDNA under control of the human cytomegalovirus p
romoter but separated from it by an extraneous spacer sequence flanked
by loxP sites which blocked luciferase expression. Upon coinfection o
f 293 or MRCS cells with AdMA19 and AdCre, luciferase expression was s
pecifically induced by Cre-mediated excision of the intervening sequen
ce. The use of Ad vectors combined with the Cre-loxP system for regula
tion of gene expression and other possible applications is discussed.