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    • 1. 发明申请
    • TEMPERATURE-INDUCED POLYNUCLEOTIDES AND USES THEREFOR
    • 温度诱导的多核苷酸及其用途
    • WO2010054362A2
    • 2010-05-14
    • PCT/US2009063838
    • 2009-11-10
    • WYETH LLCMCCARTHY KEVIN MHELLER-HARRISON ROBINLEONARD MARK WCHARLEBOIS TIMOTHY S
    • MCCARTHY KEVIN MHELLER-HARRISON ROBINLEONARD MARK WCHARLEBOIS TIMOTHY S
    • C07K14/47C12N15/67C12N15/85
    • C07K14/47C12N15/63
    • The present invention is based on the discovery that the transcriptional activity of both hamster mammary tumor 7 (HMT 7) and hamster layilin genes are increased when Chinese Hamster Ovary (CHO) cells are cultured at temperatures lower than 37°C. The present invention provides novel genomic, cDNA, and protein sequences of HMT 7 and a novel genomic sequence for hamster layilin. Included in the genomic sequences are novel temperature-induced promoter sequences for each; two promoter sequences are provided for HMT 7 and one promoter sequence is provided for layilin. The invention additionally provides antisense and siRNA molecules to the nucleic acid molecules of HMT 7. The invention also provides genetically engineered expression vectors comprising the novel temperature-induced promoters of the invention, which are particularly useful as part of a mammalian temperature-inducible expression system. The invention also provides host cells and/or transgenic animals comprising the novel nucleic acid molecules of the invention. The invention further provides methods of using the polynucleotides of the invention, e.g., for temperature-induced transgene expression.
    • 本发明基于这样的发现,即当中国仓鼠卵巢(CHO)细胞在低于37℃的温度下培养时,仓鼠乳房肿瘤7(HMT 7)和仓鼠layilin基因的转录活性均增加。 本发明提供了HMT7的新型基因组,cDNA和蛋白质序列以及仓鼠layilin的新型基因组序列。 包含在基因组序列中的是各自的新型温度诱导启动子序列; 为HMT 7提供两个启动子序列,为layilin提供一个启动子序列。 本发明另外为HMT7的核酸分子提供反义和siRNA分子。本发明还提供了包含本发明的新型温度诱导启动子的基因工程表达载体,其特别用作哺乳动物温度诱导型表达系统的一部分 。 本发明还提供了包含本发明的新型核酸分子的宿主细胞和/或转基因动物。 本发明进一步提供了使用本发明的多核苷酸的方法,例如用于温度诱导的转基因表达。
    • 2. 发明申请
    • NOVEL POLYNUCLEOTIDES RELATED TO OLIGONUCLEOTIDE ARRAYS TO MONITOR GENE EXPRESSION
    • 与寡核苷酸阵列相关的新型多核苷酸监测基因表达
    • WO2006025879A3
    • 2007-01-25
    • PCT/US2005016425
    • 2005-05-11
    • WYETH CORPMELVILLE MARK WCHARLEBOIS TIMOTHY SMOUNTS WILLIAM MHANN LOUANE ESINACORE MARTIN SLEONARD MARK WBROWN EUGENE LMILLER CHRISTOPHER PLEE GENE W
    • MELVILLE MARK WCHARLEBOIS TIMOTHY SMOUNTS WILLIAM MHANN LOUANE ESINACORE MARTIN SLEONARD MARK WBROWN EUGENE LMILLER CHRISTOPHER PLEE GENE W
    • C12Q1/68G06F19/00
    • C07K14/47C12Q1/6837C12Q1/6876C12Q2600/158
    • The present invention provides an oligonucleotide array capable of identifying genes and related pathways involved with the induction of a particular phenotype by a cell line, e.g., the genes and related pathways involved with the induction of transgene expression by the cell line. The invention is particularly useful when there is a little or no information about the genome of the cell line being studied, because it provides methods for identifying consensus sequences for known and previously undiscovered genes, and for designing oligonucleotide probes to the identified consensus sequences. Additionally, when the array is to be used to determine optimal conditions for expression of a transgene by the cell line, the invention teaches methods of including oligonucleotide probes to transgene sequences in the array. The invention also provides methods of using the array to identify genes and related pathways involved with the induction of a particular cell line phenotype. The invention also provides novel polynucleotides of undiscovered genes (i.e., a gene that had not been sequenced and/or shown to be expressed by CHO cells) and novel polynucleotides involved with the induction of a particular cell phenotype, e.g., increased survival when grown under stressful culture conditions, increased transgene expression, decreased production of an antigen, etc. These novel polynucleotides are termed novel CHO sequences and differential CHO sequences, respectively. The invention also provides genetically engineered expression vectors, host cells, and transgenic animals comprising the novel nucleic acid molecules of the invention. The invention additionally provides antisense and RNAi molecules to the nucleic acid molecules of the invention. The invention further provides methods of using the polynucleotides of the invention.
    • 本发明提供了能够鉴定涉及通过细胞系诱导特定表型的基因和相关途径的寡核苷酸阵列,例如涉及通过细胞系诱导转基因表达的基因和相关途径。 当有关于正在研究的细胞系的基因组的信息很少或没有信息时,本发明是特别有用的,因为它提供了用于鉴定已知和先前未被发现的基因的共有序列的方法,以及为鉴定的共有序列设计寡核苷酸探针。 另外,当该阵列用于确定由细胞系表达转基因的最佳条件时,本发明教导了将寡核苷酸探针包括在阵列中的转基因序列的方法。 本发明还提供了使用该阵列鉴定涉及诱导特定细胞系表型的基因和相关途径的方法。 本发明还提供未被发现的基因的新型多核苷酸(即,未被测序和/或显示由CHO细胞表达的基因)和参与诱导特定细胞表型的新型多核苷酸,例如当在下面生长时增加的存活 压力培养条件,转基因表达增加,抗原产生减少等。这些新型多核苷酸分别被称为新的CHO序列和差异CHO序列。 本发明还提供基因工程表达载体,宿主细胞和包含本发明的新型核酸分子的转基因动物。 本发明还向本发明的核酸分子提供反义和RNAi分子。 本发明还提供使用本发明的多核苷酸的方法。