FLP-mediated gene modification in mammalian cells, and compositions and cells useful therefor
First Claim
1. A method for excising a nucleic acid sequence from the genome of a mammalian host cell, wherein said nucleic acid sequence is located in the genome of said mammalian host cell between a first FLP recombination target site (FRT) and a second FRT, said method comprising introducing into said mammalian host cell an FLP recombinase, wherein said FLP recombinase catalyzes recombination between said first FRT and said second FRT, thereby excising said nucleic acid sequence from said genome.
0 Assignments
0 Petitions
Accused Products
Abstract
A gene activation/inactivation and site-specific integration system has been developed for mammalian cells. The invention system is based on the recombination of transfected sequences by FLP, a recombinase derived from Saccharomyces. In several cell lines, FLP has been shown to rapidly and precisely recombine copies of its specific target sequence. For example, a chromosomally integrated, silent β-galactosidase reporter gene was activated for expression by FLP-mediated removal of intervening sequences to generate clones of marked cells. Alternatively, the reverse reaction can be used to target transfected DNA to specific chromosomal sites. These results demonstrate that FLP can be used, for example, to mosaically activate or inactivate transgenes for a variety of therapeutic purposes, as well as for analysis of vertebriate development.
-
Citations
25 Claims
- 1. A method for excising a nucleic acid sequence from the genome of a mammalian host cell, wherein said nucleic acid sequence is located in the genome of said mammalian host cell between a first FLP recombination target site (FRT) and a second FRT, said method comprising introducing into said mammalian host cell an FLP recombinase, wherein said FLP recombinase catalyzes recombination between said first FRT and said second FRT, thereby excising said nucleic acid sequence from said genome.
-
18. A method for the recovery of an isolated nucleic acid sequence from the genome of a mammalian host cell, wherein said nucleic acid sequence is bordered on the 5'"'"' and 3'"'"' ends with a FRT, said method comprising:
-
a) contacting genomic DNA from said host cell with FLP recombinase, thereby excising said nucleic acid sequence from said genome, and then b) recovering said excised nucleic acid sequence.
-
- 19. An isolated DNA construct comprising an insert segment flanked by a first FRT and a second FRT.
Specification