Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1996-01-16
1997-10-21
Ketter, James
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
4351723, 4353201, 536 232, 536 245, C12Q 168, C12N 1500, C07H 2104
Patent
active
056795238
ABSTRACT:
Methods are provided for identifying a gene at a random chromosomal locus in the genome of a mammalian cell. The method involves inactivating one copy of the gene by integrating one DNA construct (knockout construct) in that gene copy. The knockout construct includes a positive selection marker region sequence and, in a 5' direction from the selection marker region sequence, a transcription initiation region sequence responsive to a transactivation factor, said transcription initiation region oriented for antisense RNA transcription in the direction away from the selection marker region sequence. The second copy of the gene is inactivated by transforming the cells with a second DNA construct (transactivation construct) containing a gene sequence for the transactivation factor which initiates antisense RNA transcription extending from the knockout construct into the chromosomal locus flanking the knockout construct at its 5' end. Inactivation of both gene copies may result in a change in cell phenotype distinguishable from the wild-type phenotype. Optionally, the wild-type phenotype can be regained by introducing a third construct that can inhibit antisense RNA transcription.
REFERENCES:
Baim et al., "A Chimeric Mammalian Transactivator Based on the lac Repressor that is Regulated by Temperature and Isopropyl .beta.-D-thiogalactopyranoside," PNAS USA (1991), 88:5072-5076.
Lemke, "The Molecular Genetics of Myelination: An Update," GLIA (1993), 7:263-271.
Gossen et al., "Transcriptional Activation by Tetracyclines in Mammalian Cells," Science (1995), 268:1766-1769.
Stokes et al., "The Partial 3'-Conserved Segment Duplications in the Inegrons In6 from pSa and In7 from pDGO100 Have a Common Origin," Plasmid (1993), 30:39-50.
Kamano et al., "Effects of the Antisense v-myb' Expression on K562 Human Leukemia Cell Proliferation and Differentiation," Leukemia Research (1990), 14:831-839.
Baubonis and Sauer, "Genomic Targeting with Purified Cre Recombinase," Nucleic Acids Research (1993), 21:2025-2029.
Craig, "The Mechanism of Conservative Site-Specific Recombination," Annu. Rev. Genet. (1988), 22:77-105.
van der Krol et al., "Modulation of Eukaryotic Gene Expression by Complementary RNA or DNA Sequences," BioTechniques (1988), 6:958-976.
Katsuki et al., "Conversion of Normal Behavior to Shiverer by Myelin Basic Protein Antisense cDNA in Transgenic Mice," Science (1988), 593:595.
Owens and Boyd, "Expressing Antisense Po RNA in Schwann Cells Perturbs Myelination," Development (1991), 112:639-649.
Owens and Bunge, "Schwann Cells Infected with a Recombinant Retrovirus Expressing Myelin-Associated Glycoprotein Antisense RNA Do Not Form Myelin," Neuron (1991), 7:565-575.
Giese et al., "Mouse P.sub.o Gene Disruption Leads to Hypomyelination, Abnormal Expression of Recognition Molecules, and Degeneration of Myelin and Axons," Cell (1992), 71:565-576.
Cohen Stanley N.
Li Limin
Brusca John S.
Ketter James
Sherwood Pamela J.
The Board of Trustees of the Leland Stanford Junior University
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