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.
The present invention provides hyperactive piggyBac transposons, in particular hyperactive piggyBac transposons from (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.
The present invention provides hyperactive piggyBac transposons, in particular hyperactive piggyBac transposons from (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.
Gain Of Function Mutations In Atp-Dependent Transposition Proteins
Johns Hopkins University School of Medicine - Baltimore MD
International Classification:
C07K 100
US Classification:
530350, 536 232
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 Tn. Such mutations enable the insertion of a Tn 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.
Trichoplusia Ni Piggybac Transposases With Reduced Integration Activity
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.
- Baltimore MD, US Nancy L. Craig - Baltimore MD, US
International Classification:
C12N 15/90 A61K 38/45 C12N 9/12
Abstract:
The present invention provides hyperactive piggyBac transposons, in particular hyperactive piggyBac transposons from (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.
Trichoplusia Ni Piggybac Transposases With Reduced Integration Activity
HOWARD HUGUES MEDICAL INSTITUTE - Chevy Chase MD THE JOHNS HOPKINS UNIVERSITY - Baltimore MD
International Classification:
C12N 9/12 C12N 15/90
US Classification:
435462, 435194
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 The piggyBac transposases of the present invention are functionally active or hyperactive for excision and have decreased integration activity compared to wild type 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.