Lethality-based selection of recombinant genes in mammalian cells: application to identifying tumor antigens.
Many biological processes result in either cell death or cessation of cell growth. However, plasmid- and retrovirus-based mammalian expression vectors in which it has been possible to construct representative cDNA libraries cannot be readily recovered from cells that are not actively dividing. This has limited the efficiency of selection of recombinant genes that mediate either lytic events or growth arrest. Examples include genes that encode the target antigens of cytotoxic T cells, genes that promote stem-cell differentiation and pro-apoptotic genes. We have successfully constructed representative cDNA libraries in a poxvirus-based vector that can be recovered from cells that have undergone lethality-based selection. This strategy has been applied to selection of a gene that encodes a cytotoxic T-cell target antigen common to several independently derived tumors.
Smith
E S
ES
Vaccinex, Inc., Rochester, New York, USA.
Mandokhot
A
A
Evans
E E
EE
Mueller
L
L
Borrello
M A
MA
Sahasrabudhe
D M
DM
Zauderer
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics*; D001483:Base Sequence; D018076:DNA, Complementary; D005786:Gene Expression Regulation; D005804:Genes, Lethal; D005822:Genetic Vectors; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics*; D000090822:Ribosomal Protein L3; D012269:Ribosomal Proteins / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics
D000951:Antigens, Neoplasm; D018076:DNA, Complementary; D011994:Recombinant Proteins; D000090822:Ribosomal Protein L3; D012269:Ribosomal Proteins