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US 2003/0194696 A1Published application
Methods of producing a library and methods of selecting polynucleotides of interest
AssigneeUniversity of Rochester
InventorsMaurice Zauderer, Ernest S. Smith
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Abstract
The present invention relates to a high efficiency method of introducing DNA into linear DNA viruses such as poxvirus, a method of producing libraries in linear DNA viruses such as poxvirus, and methods of selecting or screening for polynucleotides of interest based on cell nonviability or other phenotypes of eukaryotic cells, especially mammalian cells.
Claims30
A method of selecting a target polynucleotide, comprising:
- (a) introducing into a population of vertebrate host cells a library of insert polynucleotides;
- wherein the host cells comprise a reporter gene encoding an endogenous protein;
- wherein the library is constructed in a linear DNA virus vector;
- wherein at least one of the insert polynucleotides comprises the target polynucleotide;
- and wherein expression of the target polynucleotide in a cell directly or indirectly alters expression of the endogenous protein;
- (b) culturing the host cells;
- and (c) collecting insert polynucleotides from those host cells which exhibit the altered expression.
4 more elements
A method of selecting a target polynucleotide, comprising:
- (a) introducing into a population of vertebrate host cells a library of insert polynucleotides;
- wherein the host cells comprise a reporter gene under operable control of a promoter which is preferentially expressed in cancerous cells or during differentiation;
- wherein the library is constructed in a linear DNA virus vector;
- wherein at least one of the insert polynucleotides comprises the target polynucleotide;
- and wherein expression of the target polynucleotide in a cell directly or indirectly alters transcription of the promoter;
- (b) culturing the host cells;
- and (c) collecting insert polynucleotides from those host cells which exhibit altered expression of the product encoded by the reporter gene.
4 more elements
A method of selecting a target polynucleotide, comprising:
- (a) introducing into a population of vertebrate host cells a library of insert polynucleotides;
- wherein said library is constructed in a linear DNA virus vector;
- wherein at least one of said insert polynucleotides comprises the target polynucleotide;
- and wherein expression of said target polynucleotide directly or indirectly alters a phenotype in a cell comprising said target polynucleotide;
- (b) culturing said host cells;
- and (c) collecting insert polynucleotides from those host cells which exhibit said altered phenotype.
3 more elements
Dependencies are inferred from the claim text.