Multicellular living organisms and unmodified parts thereof and – Nonhuman animal – Transgenic nonhuman animal
Patent
1995-02-24
1999-06-29
Chambers, Jasemine C.
Multicellular living organisms and unmodified parts thereof and
Nonhuman animal
Transgenic nonhuman animal
435 6, 435 691, 435 701, 435 703, 435 914, 435455, 435462, 435463, 435325, 4353201, 536 241, C12N 500, C12N 1500, C12N 1509, C12N 1563
Patent
active
059171227
DESCRIPTION:
BRIEF SUMMARY
1. INTRODUCTION
The present invention relates to a tetracycline repressor-mediated binary regulation system for the control of gene expression in transgenic animals. It is based, at least in part, on the discovery that, in a non-human transgenic animal that carries a first transgene under the control of a modified promoter comprising a tetR operator sequence and a second transgene encoding the tetR repressor protein, expression of the first transgene may be efficiently induced by administering tetracycline to the animal.
2. BACKGROUND OF THE INVENTION
2.1. Control of Gene Expression in Transgenic Animals
The production of transgenic animals for both experiment and agricultural purposes is now well known (Wilmut et al., Jul. 7, 1988, New Scientist pp. 56-59). In research, transgenic animals are a powerful tool that have made significant contributions to our understanding of many aspects of biology and have contributed to the development of animal models for human diseases (Jaenisch, 1988, Science 240:1468-1474). It is also clear that several livestock species can be made transgenic and these species promise to expand and revolutionize the method of production and diversity of pharmaceutical products available in the future, in addition to improving the agricultural qualities of the livestock species (Wilmut et al., supra).
A critical, often neglected, aspect of developing transgenic animals is the process whereby expression of the newly introduced gene, referred to as the transgene, is controlled. This is an important process since stringent regulation of transgene expression is often important both for practical, regulatory and safety reasons and to maintain the health of the transgenic animal. In the past either "inducible" or "tissue specific" regulatory mechanisms have been used. Inducible regulation is defined herein as a method of gene regulation which allows for some form of outside manipulation of the onset and/or level of transgene expression. Tissue specific regulation is defined herein as a method for targeting transgene expression to particular tissues or organs.
Inducible gene regulation may be achieved using relatively simple promoter systems such as the metallothionein heat shock promoters, or by using promoters which are responsive to specific compounds such as the Mouse mammary tumor virus LTR which is responsive to glucocorticoid stimulation. More flexible, though more complex inducible regulation systems can be achieved through a "binary" gene approach which utilizes a transactivator gene product to control expression of a second gene of interest. Tissue specific gene regulation usually consists of simple single gene methods (Byrne et al., 1989, Proc. Natl. Acad. Sci. U.S.A. 86:5473-5477; Ornitz et al., 1991, Proc. Natl. Acad. Sci. U.S.A. 88:698-702), although binary transactivator systems can also provide a high degree of tissue specificity.
These current systems provide only a limited ability to control the time of transgene expression within individual animals. In this respect tissue specific promoter elements provide no method to control the onset of transgene activity, but function merely to target gene expression to defined sites. Simple inducible promoters such as metallothionein generally lack tissue specificity and usually have some aspect of endogenous basal expression which cannot be controlled. Thus even for the extensively used inducible metallothionein promoter this approach at best only permits selection of the time at which a relative increase in transgene expression can be induced.
Binary transactivation systems typically consist of two transgenic animals. One animal contains the gene of interest controlled by a promoter element that requires a specific transactivator gene product for expression. Thus, the gene of interest is not expressed in the absence of the transactivator. A second transgenic animal is then made which expresses the required transactivator in the desired tissue. By mating these two transgenic animals, offspring containing both the gene of intere
REFERENCES:
patent: 5075229 (1991-12-01), Hanson et al.
patent: 5221778 (1993-06-01), Byrne et al.
Byrne and Ruddle, 1989, "Multiplex gene regulation: A two-tiered aproach to transgene regulation in transgenic mice", Proc Natl Acad Sci 86:5473-5477.
Gatz et al., 1991, "Regulation of a modified CaMV 35S promoter by the Tn10-encoded Tet repressor in transgenic tobacco", Mol Gen Genet 227:229-237.
Gatz and Quail, 1988, "Tn10-encoded tet repressor can regulate an operator-containing plant promoter", Proc Natl Acad Sci 85:1394-1397.
Gatz et al., 1992, "Stringent repression and homogeneous de-repression bytetracycline of a modified CaMV 35S promoter in intact transgenic tobacco plants", Plant J 2(3):397-404.
Gossen and Bujard, 1992, "Tight control of gene expression in mammalian cells by tetracycline-responsive promoters", Proc Natl Acad Sci 89:5547-5551.
Khillan et al., 1988, "Gene transcativation mediated by the TAT gene of human immunodeficiency virus in transgenic mice", Nucl Acids Res 16(4):1423-1430.
Ornitz et al., 1991, "Binary system for regulatin transgene expression in mice: Targeting int-2 gene expression with yeast GAL4/UAS control elements", Proc Natl Acad Sci 88:698-702.
Wilde et al., 1992, "Control of gene expression in tobacco cells using a bacterial operator-repressor system", EMBO J 11(4):1251-1259.
P. Beufey et al. Embo J. 9(6) 1677-84, 1990.
J. Watson et al., Molecular Biol. of the Gene, 4.sup.th Ed., Benjamin/Cummings Publ. Co., Inc., (87), Menlo Park, CA, pp. 252-253 & 440-442.
T. Giordano et al., Gene, (90) 88(2): 288-8.
Chambers Jasemine C.
Martin Jill D.
LandOfFree
Tetracycline repressor-mediated binary regulation system for con does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Tetracycline repressor-mediated binary regulation system for con, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Tetracycline repressor-mediated binary regulation system for con will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1377015