Back to results
US 6,080,576 AGranted
Vectors for gene trapping and gene activation
AssigneeLexicon Genetics Incorporated
InventorsBrian Zambrowicz, Glenn Friedrich, Arthur T. Sands
Life of this patent
Estimating the term…
Abstract
A novel 3' gene trap cassette is described that does not encode a marker conferring antibiotic resistance and can be used to efficiently trap and identify cellular genes. Vectors incorporating the presently 3' gene trap cassette find particular application in gene discovery, the production of transgenic cells and animals, and gene activation.
Claims21
A genetically engineered vector comprising:
- a) a 5' gene trap cassette comprising in operable combination:
- 1) a splice acceptor;
- 2) a first exon sequence located 3' to said splice acceptor, said first exon encoding a marker enabling the identification of a cell expressing said exon;
- and
- 3) a polyadenylation sequence defining the 3' end of said first exon;
- b) a 3' gene trap cassette located 3' to said polyadenylation sequence comprising in operable combination:
- 1) a first promoter;
- 2) a second exon sequence located 3' from and expressed by said promoter, said second exon being derived from a naturally occuring eukaryotic gene, said second exon not encoding an activity conferring antibiotic resistance and said second exon not being a reporter gene;
- 3) a splice donor sequence defining the 3' region of the exon said splice donor sequence being derived from a naturally occuring eukaryotic gene;
- and
- wherein said vector docs not encode a promoter mediating the expression of said first exon, and wherein said vector does not encode a sequence that mediates the polyadenylation of an mRNA transcript encoded by said second exon sequence and expressed by said first promoter.
8 more elements
A genetically engineered vector comprising:
- a) a marker gene expressed by a vector encoded promoter;
- and
- b) a 3' gene trap cassette comprising in operable combination:
- 1) a second promoter;
- 2) an exon sequence located 3' from and expressed by said second promoters said exon being derived from a naturally occuring, eukaryotic gene, said second exon not encoding an activity conferring antibiotic resistance and said exon not being a reporter gene;
- 3) a splice donor sequence defining the 3' region of the exon, said splice donor sequence being derived from a naturally occuring eukaryotic acne;
- and
- wherein said vector does not encode a sequence that mediates the polyadenylation of an mRNA transcript encoded by said exon sequence.
5 more elements
A genetically engineered retroviral vector comprising:
- (a) a first retroviral LTR sequence;
- (b) a promoter operative in eukaryotic cells;
- (c) an exon sequence located 3' from and expressed by said promoter, said exon being derived from a naturally occuring eukaryotic gene, said second exon not encoding an activity conferring antibiotic resistance and said exon not being a reporter gene;
- (d) a splice donor sequence defining the 3' region of the exon said splice donor sequence being derived from a naturally occuring eukaryotic gene;
- and
- (e) a second retroviral LTR sequence;
- wherein said promoter, exon and splice donor are present in the vector in between said first and second LTR sequence and in an opposite orientation to said first and second retroviral LTR sequences and wherein said vector does not incorporate a sequence that mediates the polyadenylation of an mRNA transcript expressed by said promoter of element (b) and encoded by said exon of element (c).
4 more elements
A method to alter the expression of a gene in an isolated eukaryotic cell comprising introducing a 3' gene trap cassette vector into said cell, said 3' gene trap cassette comprising in operable combination:
- 1) a promoter;
- 2) an exon sequence located 3' from and expressed by said promoter, said exon being derived from a naturally occuring eukaryoric one said exon not encoding an activity conferring antibiotic resistance and said exon not being a reporter gene;
- and
- 3) a splice donor sequence defining the 3' region of said exon said slice donor sequence being derived from a naturally occuring eukaryotic gene;
- wherein said cassette is non-homologously incorporated into the genome of an eukaryotic target cell and said splice donor sequence of the transcript encoded by said exon is spliced to a splice acceptor sequence of said cellularly encoded gene.
2 more elements
Dependencies are inferred from the claim text.