Method of eliminating inhibitory/ instability regions of mRNA
Patent 5965726 Issued on October 12, 1999. Estimated Expiration Date: May 2, 2017. Estimated Expiration Date is calculated based on simple USPTO term provisions. It does not account for terminal disclaimers, term adjustments, failure to pay maintenance fees, or other factors which might affect the term of a patent.
A method of locating an inhibitory/instability sequence or sequences within the coding region of an mRNA and modifying the gene encoding that mRNA to remove these inhibitory/instability sequences by making clustered nucleotide substitutions without altering the coding capacity of the gene is disclosed. Constructs containing these mutated genes and host cells containing these constructs are also disclosed. The method and constructs are exemplified by the mutation of a Human Immunodeficiency Virus-1 Rev-dependent gag gene to a Rev-independent gag gene. Constructs useful in locating inhibitory/instability sequences within either the coding region or the 3' untranslated region of an mRNA are also disclosed. The exemplified constructs of the invention may also be useful in HIV-1 immunotherapy and immunoprophylaxis.
Other References
B.A. Bunnell et al., "A Dominant Negative Mutation in Two Proteins Created by Ectopic Expression of an Au-Rich 3' Untranslated Region", Somataic Cell and Mol. Genet. 16:151-162 (1990)
D. Caput et al., "Identification of a common nucleotide sequence in the 3'-untranslated region of mRNA molecules specifying inflammatory mediators", Proc. Natl. Acad. Sci. 83:1670-1674 (1986)
P. Carter-Muenchau and R. Wolf, "Growth-rate-dependent regulation of 6-phosphogluconate dehydrogenase level mediated by an anti-Shine-Dalgarno sequence located within the Escherichia coli gnd structural gene", Proc. Natl. Acad. Sci., USA, 86:1138-1142 (1989)
A.W. Cochrane et al., "Identification and Characterization of Intragenic Sequences Which Repress Human Immunodeficiency Virus Structural Gene Expression", J. Virol. 65:5303-5313 (1991)
M.D. Cole and S.E. Mango, cis-Acting Determinants of c-myc mRNA Stability, Enzyme 44:167-180 (1990)
M.D. Edge et al. "Total synthesis of a human leukocyte interferone gene", Nature 292:756-762 (1981)
M. Emerman, "The rev Gene Product of the Human Immunodeficiency Virus Affects Envelope-Specific RNA Localization", Cell 57:1155-1165 (1989)
B. Felber et al., "rev protein of human immunodeficiency virus type 1 affects the stability and transport of the viral mRNA", Proc. Natl. Acad. Sci. USA 86:1495-1499 (1989)
M. Hadzopoulou-Cladaras et al., "The rev (trs/art) Protein of Human Immunodeficiency Virus Type 1 Affects Viral mRNA and Protein Expression via a cis-Acting Sequence in the env Region", J. Virol. 63:1265-1274 (1989)
M.W. Hentze, "Determinants and regulation of cytoplasmic mRNA stability in eukaryotic cells", Biochem. Biophys. Acta 1090: 281-292 (1991)
E. Jay et al., "Chemical Synthesis of a Biologically Active Gene for Human Immune Interferon-γ", J. Biol. Chem. 259:6311-6317 (1984)
T. R. Jones and M.D. Cole, "Rapid Cytoplasmic Turnover of c-myc mRNA: Requirement of the 3' Untranslated Sequences", Mol. Cell Biol. 7:4513-4521 (1987)
R. Kamen et al., "A Novel Mechanism of Post Transcriptional, Sequence-Specific Regulation of mRNA Stability", J. Cell Bio. Supp. 10D (1986):152 (Abst. No. 0210)
D.M. Koeller et al., "Translation and the stability of mRNAs encoding the transferrin receptor and c-fos", Proc. Natl. Acad. Sci. USA, 88:7778-7782 (1991)
V. Kruys et al., "Constitutive activity of the tumor necrosis factor promoter is canceled by the 3' untranslated region in nonmacrophage cell lines; a trans-dominant factor overcomes this suppressive effect", Proc. Natl. Acad. Sci. USA, 89:673-677 (1992)
T.A. Kunkel, "Rapid and efficient site-specific mutagenesis without phenotypic selection", Proc. Natl. Acad. Sci. USA, 82:488-492 (1985)
I.A. Laird-Offringa et al., "Rapid c-myc mRNA degradation does not require (A+U)-rich sequences or complete translation of the mRNA", Nucleic Acids Res. 19:2387-2394 (1991)
M.D. Lundigran et al., "Transcribed sequences of the Escherichia coli btuB gene control its expression and regulation by vitamin B12 ", Proc. Natl. Acad. Sci. USA, 88:1479-1483 (1991)
F. Maldarelli et al., "Identification of Posttranscriptionally Active Inhibitory Sequences in Human Immunodeficiency Virus Type 1 RNA: Novel Level of Gene Regulation", J. Virol. 65:5732-5743 (1991)
K.P. Nambiar et al., "Total synthesis and Cloning of a Gene Coding for the Ribonuclease S Protein", Science 223:1299-1301 (1984)
R. Parker and A. Jacobson, "Translation and a 42-nucleotide segment within the coding region of the mRNA encoded by the MATl gene are involved in promoting rapid mRNA decay in yeast", Proc. Natl. Acad. Sci. USA, 87:2780-2784 (1990)
C.A. Rosen, "Intragenic cis-acting art gene-responsive sequences of the human immunodeficiency virus", Proc. Natl. Acad. Sci., USA, 85:2071-2075 (1988)
S. Schwartz et al., "Distinct RNA Sequences in the gag region of Human Immunodeficiency Virus Type 1 Decrease RNA Stability and Inhibit Expression in the Absence of Rev Protein", J. Virol. 66:150-159 (1992)
G. Shaw and R. Kamen, "A Conserved AU Sequence from the 3' Untranslated Region of GM-CSF mRNA Mediates Selective mRNA Degradation", Cell 46:659-667 (1986)
G. Shaw and R. Kamen, "A Conserved AU Sequence from the 3' Untranslated Region of GM-CSF mRNA Mediates Selective mRNA Degradation", J. Cell. Bio. Supp. 11C (1987):132 (Abst. No. L541)
A.-B. Shyu et al., "Two distinct destabilizing elements in the c-fos message trigger deadenylation as a first step in rapid mRNA decay", Gen. & Devel. 5:221-231 (1991)
C.M. Stoltzfus and S.J. Fogarty, "Multiple Regions in the Rous Sarcoma Virus src Gene Intron Act in cis To Affect the Accumulation of Unspliced RNA", J. Virol. 63:1669-1676 (1989)
T. Wilson and R. Treisman, "Removal of poly(A) and consequent degradation of c-fos mRNA facilitated by 3' AU-rich sequences", Nature 336:396-399 (1988)
R. Wisdom and W. Lee, "The protein-coding region of c-myc mRNA contains a sequence that specifies rapid mRNA turnover and induction by protein synthesis inhibitors", Gen. & Devel. 5:232-243 (1991)
D.H. Wreschner and G. Rechavi, "Differential mRNA stability to reticulocyte ribonucleases correlates with 3' non-coding (U)n A sequences", Eur. J. Biochem. 172:333-340 (1988)
Schwartz et al., "Mutational Inactivation of an Inhibitory Sequence in HIV-1 Results in Rev-independent gag Expression", Journal of Virology, vol. 66, 12:7176-7182 (1992) (Not prior art: published after filing date of 07/858,747 application.)
Copy of Search Report
Schwartz, et al, "Mutational Inactivation of an . . . " J. Virol. 66(12):7176-7182, 1992
Haynes, B.F. "Scientific and Social Issues of . . . " Science vol. 260, pp. 1279-1286, 1993
Butisi, et al, "Comparative Analysis of HIV-Specific . . . " Abstract J306, J. Cell. Biochem. Suppl. 18B, 1994
Fox, No Winners Against AIDS. Bio/Technology vol. 12:128, 199