Abstract: The present invention provides hyperactive piggyBac transposons, in particular hyperactive piggyBac transposons from Trichoplusia ni (cabbage looper moth) that transpose at a higher frequency than wildtype. The invention also features integration defective piggyBac transposons. The piggyBac transposons and transposases can be used in gene transfer systems for stably introducing nucleic acids into the DNA of a cell. The gene transfer system can be used in methods, for example, but not limited to, gene therapy, insertional mutagenesis, or gene discovery.

Abstract: The present invention is directed to nucleic acid and amino acid sequences of a novel piggyBac transposase enzymes created by modifying the transposase of Trichoplusia ni. The piggyBac transposases of the present invention are functionally active or hyperactive for excision and have decreased integration activity compared to wild type Trichoplusia ni piggyBac transposase enzyme. These transposases are ideal for use in methods of transforming cells and organisms. In particular embodiments, the present invention provides methods of transient integration and expression of transgenes.

Abstract: The present invention is directed to nucleic acid and amino acid sequences of a novel piggyBac transposase enzymes created by modifying the transposase of Trichoplusia ni. The piggyBac transposases of the present invention are functionally active or hyperactive for excision and have decreased integration activity compared to wild type Trichoplusia ni piggyBac transposase enzyme. These transposases are ideal for use in methods of transforming cells and organisms. In particular embodiments, the present invention provides methods of transient integration and expression of transgenes.

Abstract: The present invention is directed to nucleic acid and amino acid sequences of a novel piggyBac transposase enzymes created by modifying the transposase of Trichoplusia ni. The piggyBac transposases of the present invention are functionally active or hyperactive for excision and have decreased integration activity compared to wild type Trichoplusia ni piggyBac transposase enzyme. These transposases are ideal for use in methods of transforming cells and organisms. In particular embodiments, the present invention provides methods of transient integration and expression of transgenes.

Abstract: The present invention provides hyperactive piggyBac transposons, in particular hyperactive piggyBac transposons from Trichoplusia ni (cabbage looper moth) that transpose at a higher frequency than wildtype. The invention also features integration defective piggyBac transposons. The piggyBac transposons and transposases can be used in gene transfer systems for stably introducing nucleic acids into the DNA of a cell. The gene transfer system can be used in methods, for example, but not limited to, gene therapy, insertional mutagenesis, or gene discovery.

Abstract: Hyperactive Hermes Transposase mutants and genes encoding them are disclosed. These transposases are easily purified in large quantity after expression in bacteria. The modified Hermes Transposases are soluble and stable and exist as smaller active complexes compared to the native enzyme. The consensus target DNA recognition sequence is the same as the native enzyme and shows minimal insertional sequence bias. These properties are useful in whole genome sequencing applications that involve sample DNA preparation requiring simultaneous fragmentation and attachment of custom sequences to the ends of the fragments. Methods and compositions using these transposases in fragmentation and 5? end-tagging are also disclosed.

Abstract: The present invention is directed to nucleic acid and amino acid sequences of a novel piggyBac transposase enzymes created by modifying the transposase of Trichoplusia ni. The piggyBac transposases of the present invention are functionally active or hyperactive for excision and have decreased integration activity compared to wild type Trichoplusia ni piggyBac transposase enzyme. These transposases are ideal for use in methods of transforming cells and organisms. In particular embodiments, the present invention provides methods of transient integration and expression of transgenes.

Abstract: The invention is specifically directed to efficient, random, simple insertion of a transposon or derivative transposable element into DNA in vivo or in vitro. The invention is particularly directed to mutations in ATP-utilizing regulatory transposition proteins that permit insertion with less target-site specificity than wild-type. The invention encompasses gain-of-function mutations in TnsC, an ATP-utilizing regulatory transposition protein that activates the bacterial transposon Tn7. Such mutations enable the insertion of a Tn7 transposon or derivative transposable element in a non-specific manner into a given DNA segment. Insertion can be effected in plasmid and cosmid libraries, cDNA libraries, PCR products, bacterial artificial chromosomes, yeast artificial chromosomes, mammalian artificial chromosomes, genomic DNAs, and the like. Such insertion is useful in DNA sequencing methods, for genetic analysis by insertional mutagenesis, and alteration of gene expression by insertion of a given genetic sequence.

Abstract: The invention is specifically directed to efficient, random, simple insertion of a transposon or derivative transposable element into DNA in vivo or in vitro. The invention is particularly directed to mutations in ATP-utilizing regulatory transposition proteins that permit insertion with less target-site specificity than wild-type. The invention encompasses gain-of-function mutations in TnsC, an ATP-utilizing regulatory transposition protein that activates the bacterial transposon Tn7. Such mutations enable the insertion of a Tn7 transposon or derivative transposable element in a non-specific manner into a given DNA segment. Insertion can be effected in plasmid and cosmid libraries, cDNA libraries, PCR products, bacterial artificial chromosomes, yeast artificial chromosomes, mammalian artificial chromosomes, genomic DNAs, and the like. Such insertion is useful in DNA sequencing methods, for genetic analysis by insertional mutagenesis, and alteration of gene expression by insertion of a given genetic sequence.