Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification – Introduction of a polynucleotide molecule into or...
Reexamination Certificate
2001-08-09
2002-11-05
Yucel, Remy (Department: 1636)
Chemistry: molecular biology and microbiology
Process of mutation, cell fusion, or genetic modification
Introduction of a polynucleotide molecule into or...
C435S320100, C435S325000, C435S468000, C435S410000, C536S023100, C536S023200
Reexamination Certificate
active
06475798
ABSTRACT:
INTRODUCTION
1. Field of the Invention
The field of this invention is nucleic acid vectors.
2. Background of the Invention
The introduction of an exogenous nucleic acid sequence (e.g. DNA) into a cell, a process known as “transformation,” plays a major role in a variety of biotechnology and related applications, including research, synthetic and therapeutic applications. Research applications in which transformation plays a critical role include the production of transgenic cells and animals. Synthetic applications in which transformation plays a critical role include the production of peptides and proteins. Therapeutic applications in which transformation plays a key role include gene therapy applications. Because of the prevalent role transformation plays in the above and other applications, a variety of different transformation protocols have been developed.
In many transformation applications, it is desirable to introduce the exogenous DNA in a manner such that it is incorporated into a target cell's genome. One means of providing for genome integration is to employ a vector that is capable of homologous recombination. Techniques that rely on homologous recombination can be disadvantageous in that the necessary homologies may not always exist; the recombination events may be slow, etc. As such, homologous recombination based protocols are not entirely satisfactory.
Accordingly, alternative viral based transformation protocols have been developed, in which a viral vector is employed to introduce exogenous DNA into a cell and then subsequently integrate the introduced DNA into the target cell's genome. Viral based vectors finding use include retroviral vectors, e.g. Maloney murine leukemia viral based vectors. Other viral based vectors that find use include adenovirus derived vectors, HSV derived vectors, sindbis derived vectors, etc. While viral vectors provide for a number of advantages, their use is not optimal in many situations. Disadvantages associated with viral based vectors including immunogenicity, viral based complications, and the like.
Accordingly, there is continued interest in the development of additional vectors for use in transformation protocols. Of particular interest is the development of non-viral vectors that provide for stable integration of exogenous DNA in a cell genome through a mechanism other than homologous recombination.
Relevant Literature
U.S. patents of interest include: U.S. Pat. Nos. 5,719,055 and 4,670,388. Other references of interest include: Rio et al., “Identification and Immunochemical Analysis of Biologically Active Drosophila P Element Transposase,” Cell (Jan. 17, 1986) 44:21-32; and Rio et al., “Evidence for Drosophila P Element Transposase Activity in Mammalian Cells and Yeast,” J. Mol. Biol. (1988) 200: 411-415.
Additional articles of interest include: Schouten et al., Nuc. Acids Res. (1998) 26:3013-3017; Ivics et al., Cell (1997) 91: 501-510; Luo et al., Proc. Nat'l Acad. Sci USA (1998) 95:10769-10773; and Ivics et al., Proc. Nat'l Acad. Sci. USA (1996) 93:5008-5013.
SUMMARY OF THE INVENTION
P element derived vectors and methods for their use in the insertion of an exogenous nucleic acid into a target cell genome are provided. The vectors of the subject invention include a pair of P element transposase recognized insertion sequences, e.g. P element derived 31 base pair inverted repeats, flanking at least two transcriptionally active genes. In practicing the subject methods, a vector as described above carrying an exogenous nucleic acid is introduced into a target cell under conditions sufficient for transposition of the exogenous nucleic acid from the vector into the target cell genome. The subject methods find use in a variety of transformation applications, including research, polypeptide synthesis and therapeutic applications.
REFERENCES:
patent: 4670388 (1987-06-01), Rubin et al.
patent: 5719055 (1998-02-01), Cooper
patent: 6291243 (2001-09-01), Fogarty et al.
Rio et al. Evidence for Deosophila P Element Transposase Activity in Mammalian cells and yeast J. Mol. Riol. 1988 200 411-415.*
Beall et al. Drosophila P-element transposase is a novel site-specific endonuclease Genes & Development 11:2137-2151 1997.*
Ivics et al. (1996). “Identification of functional domains and evolution of Tc1-like transportable elements”Proc. Natl. Acad. Sci.,vol. 93:5008-5013.
Ivics et al. (1997). “Molecular Reconstruction of Sleeping Beauty, a Tc1-like Transposon from Fish, and its Transposition in Human Cells”Cell,vol. 91:501-510.
Luo et al. (1998), “Chromosomal Transposition of a Tc1/Mariner-like Element in Mouse Embryonic Stem Cells,”Proc. Natl. Acad. Sci. USA,vol. 95:10769-10773.
Rio et al. (1986), “Identification and Immunochemical Analysis of Biologically Active Drosophila P Element Transposase”,Cell,vol. 44:21-32.
Rio, D.C. “Regulation of Drosophilia P element transposition”,Trends in Genetics,Sep. 1991, vol. 7, No. 9, pp. 282-287.
Schouten et al. (1998), “Transposon Tc1 of the NematodeCaenorhabditis ElegansJumps in Human Cells,”Nucleic Acid Research,vol. 26(12):3013-3017.
Thummel et al., “Vectors for Drosophilia P-element-mediated transformation and tissue culture transfection”Gene, 74 (1988) 445-456.
Segal et al., “Genetic Transformation of Drosophilia Cells in Culture by P-Element-Mediated Transposition”Somatic cell and Molecular Genetics, vol. 22, No. 2, 1996, pp. 159-165.
Hay et al., “P element insertion-dependent gene activation in the Drosophilia eye”Proc. Natl. Acad. Sci. USA, vol. 94, pp. 5195-5200, May 1997.
Fogarty Patrick
Lipsick Joseph
Bozicevic, Field & Francis
Field Bret E.
Katcheves Konstantina
The Board of Trustees of the Leland Stanford Junior University
Yucel Remy
LandOfFree
P element derived vector and methods for its use does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with P element derived vector and methods for its use, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and P element derived vector and methods for its use will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2939818