In vivo genome-wide mutagenesis
Disclosed herein are compositions and methods for deleting or duplicating DNA in a mammalian genome. Also disclosed are compositions and methods for generating a random genome-wide chromosome rearrangement. Also disclosed are compositions and methods for streamlined construction of gene targeting vectors.
1. A gene-trap vector, comprising the formula:
3′TR-lox-SA-R 1 —X—R 2 -lox-X-5′TR
wherein 3′TR and 5′TR are piggyBac 3′ and 5′ transposon terminal sequences comprising 3′ and 5′ inverted terminal repeats (ITR), respectively;
wherein the lox sites are in the same orientation;
wherein SA is a splice acceptor;
wherein R 1 is a first reporter sequence linked to SA; and
wherein R 2 is a second reporter sequence functionally linked to an expression control sequence, and wherein X is a recombination site, wherein X is FRT, attB or attP.
2. A gene-trap vector, comprising the formula:
3′TR-lox-SA-R 1 —X—R 2 —X-lox-5′TR
wherein 3′TR and 5′TR are piggyBac 3′ and 5′ transposon terminal sequences comprising 3′ and 5′ inverted terminal repeats (ITR), respectively;
wherein the lox sites are in the same orientation;
wherein SA is a splice acceptor;
wherein R 1 is a first reporter sequence linked to SA; and
wherein R 2 is a second reporter sequence functionally linked to an expression control sequence, and wherein X is a recombination site, wherein X is FRT, attB or attP.
3. A gene-trap vector comprising the nucleic acid sequence set forth in SEQ ID NO:105, SEQ ID NO:106, or SEQ ID NO:107.