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    • 6. 发明申请
    • OLIGONUCLEOTIDE ARRAYS TO MONITOR GENE EXPRESSION AND METHODS FOR MAKING AND USING SAME
    • 寡核苷酸阵列监测基因表达及其制备和使用方法
    • WO2005111246A1
    • 2005-11-24
    • PCT/US2005/016880
    • 2005-05-11
    • WYETHMELVILLE, Mark, W.CHARLEBOIS, Timothy, S.MOUNTS, William, M.HANN, Louane, E.SINACORE, Martin, S.LEONARD, Mark, W.BROWN, Eugene, L.MILLER, Christopher, P.LEE, Gene, W.
    • MELVILLE, Mark, W.CHARLEBOIS, Timothy, S.MOUNTS, William, M.HANN, Louane, E.SINACORE, Martin, S.LEONARD, Mark, W.BROWN, Eugene, L.MILLER, Christopher, P.LEE, Gene, W.
    • C12Q1/68
    • 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 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,以及为所识别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。