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    • 2. 发明授权
    • Expression monitoring by hybridization to high density nucleic acid arrays
    • 通过与高密度核酸阵列杂交进行表达监测
    • US06410229B1
    • 2002-06-25
    • US09212004
    • 1998-12-14
    • David J. LockhartEugene L. BrownGordon G. WongMark S. CheeThomas R. Gingeras
    • David J. LockhartEugene L. BrownGordon G. WongMark S. CheeThomas R. Gingeras
    • C12Q168
    • C12Q1/6809C12Q1/6837G01N15/1475C12Q2565/507
    • This invention provides methods of monitoring the expression levels of a multiplicity of genes. The methods involve hybridizing a nucleic acid sample to a high density array of oligonucleotide probes where the high density array contains oligonucleotide probes complementary to subsequences of target nucleic acids in the nucleic acid sample. In one embodiment, the method involves providing a pool of target nucleic acids comprising RNA transcripts of one or more target genes, or nucleic acids derived from the RNA transcripts, hybridizing said pool of nucleic acids to an array of oligonucleotide probes immobilized on surface, where the array comprising more than 100 different oligonucleotides and each different oligonucleotide is localized in a predetermined region of the surface, the density of the different oligonucleotides is greater than about 60 different oligonucleotides per 1 cm2, and the olignucleotide probes are complementary to the RNA transcripts or nucleic acids derived from the RNA transcripts; and quantifying the hybridized nucleic acids in the array.
    • 本发明提供监测多种基因的表达水平的方法。 所述方法包括将核酸样品与寡核苷酸探针的高密度阵列杂交,其中高密度阵列含有与核酸样品中靶核酸的子序列互补的寡核苷酸探针。 在一个实施方案中,所述方法包括提供包含一个或多个靶基因的RNA转录物或衍生自RNA转录物的核酸的靶核酸库,将所述核酸库与固定在表面上的寡核苷酸探针阵列杂交,其中 包含超过100种不同寡核苷酸的阵列和每种不同的寡核苷酸被定位在表面的预定区域中,不同寡核苷酸的密度大于每1cm 2大约60个不同的寡核苷酸,并且寡核苷酸探针与RNA转录物互补 衍生自RNA转录物的核酸; 并定量阵列中的杂交核酸。
    • 4. 发明授权
    • Expression monitoring by hybridization to high density oligonucleotide arrays
    • 通过与高密度寡核苷酸阵列杂交进行表达监测
    • US06927032B2
    • 2005-08-09
    • US10353792
    • 2003-01-28
    • David J. LockhartEugene L. BrownGordon G. WongMark CheeThomas R. Gingeras
    • David J. LockhartEugene L. BrownGordon G. WongMark CheeThomas R. Gingeras
    • G01N33/53C12N15/09C12Q1/68G01N15/14G01N37/00C07H21/02C07H21/04C12M1/34C12P19/34
    • C12Q1/6809C12Q1/6837G01N15/1475C12Q2565/507
    • This invention provides methods of monitoring the expression levels of a multiplicity of genes. The methods involve hybridizing a nucleic acid sample to a high density array of oligonucleotide probes where the high density array contains oligonucleotide probes complementary to subsequences of target nucleic acids in the nucleic acid sample. In one embodiment, the method involves providing a pool of target nucleic acids comprising RNA transcripts of one or more target genes, or nucleic acids derived from the RNA transcripts, hybridizing said pool of nucleic acids to an array of oligonucleotide probes immobilized on surface, where the array comprising more than 100 different oligonucleotides and each different oligonucleotide is localized in a predetermined region of the surface, the density of the different oligonucleotides is greater than about 60 different oligonucleotides per 1 cm2, and the olignucleotide probes are complementary to the RNA transcripts or nucleic acids derived from the RNA transcripts; and quantifying the hybridized nucleic acids in the array.
    • 本发明提供监测多种基因的表达水平的方法。 所述方法包括将核酸样品与寡核苷酸探针的高密度阵列杂交,其中高密度阵列含有与核酸样品中靶核酸的子序列互补的寡核苷酸探针。 在一个实施方案中,所述方法包括提供包含一个或多个靶基因的RNA转录物或衍生自RNA转录物的核酸的靶核酸库,将所述核酸库与固定在表面上的寡核苷酸探针阵列杂交,其中 包含超过100种不同寡核苷酸的阵列和每种不同的寡核苷酸被定位在表面的预定区域中,不同寡核苷酸的密度大于每1cm 2大约60个不同的寡核苷酸,并且寡核苷酸 探针与衍生自RNA转录物的RNA转录物或核酸互补; 并定量阵列中的杂交核酸。
    • 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
    • US20100029500A1
    • 2010-02-04
    • US12492832
    • 2009-06-26
    • MARK W. MELVILLETIMOTHY S. CHARLEBOISWILLIAM M. MOUNTSLOUANE E. HANNMARTIN S. SINACOREMARK W. LEONARDEUGENE L. BROWNCHRISTOPHER P. MILLERGENE W. LEE
    • MARK W. MELVILLETIMOTHY S. CHARLEBOISWILLIAM M. MOUNTSLOUANE E. HANNMARTIN S. SINACOREMARK W. LEONARDEUGENE L. BROWNCHRISTOPHER P. MILLERGENE W. LEE
    • C40B30/04C40B40/06
    • 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,本发明教导了将阵列中的转基因序列包含寡核苷酸探针的方法。 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)和参与特定细胞表型诱导的新型多核苷酸,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。 本发明还提供了遗传工程改造的表达载体,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。
    • 7. 发明授权
    • Methods of identifying nucleic acid probes to quantify the expression of a target nucleic acid
    • 鉴定核酸探针以量化靶核酸的表达的方法
    • US06548257B2
    • 2003-04-15
    • US09935365
    • 2001-08-22
    • David J. LockhartEugene L. BrownGordon G. WongMark CheeThomas R. Gingeras
    • David J. LockhartEugene L. BrownGordon G. WongMark CheeThomas R. Gingeras
    • C12Q168
    • C12Q1/6809C12Q1/6837G01N15/1475C12Q2565/507
    • This invention provides methods of monitoring the expression levels of a multiplicity of genes. The methods involve hybridizing a nucleic acid sample to a high density array of oligonucleotide probes where the high density array contains oligonucleotide probes complementary to subsequences of target nucleic acids in the nucleic acid sample. In one embodiment, the method involves providing a pool of target nucleic acids comprising RNA transcripts of one or more target genes, or nucleic acids derived from the RNA transcripts, hybridizing said pool of nucleic acids to an array of oligonucleotide probes immobilized on surface, where the array comprising more than 100 different oligonucleotides and each different oligonucleotide is localized in a predetermined region of the surface, the density of the different oligonucleotides is greater than about 60 different oligonucleotides per 1 cm2, and the olignucleotide probes are complementary to the RNA transcripts or nucleic acids derived from the RNA transcripts; and quantifying the hybridized nucleic acids in the array.
    • 本发明提供监测多种基因的表达水平的方法。 所述方法包括将核酸样品与寡核苷酸探针的高密度阵列杂交,其中高密度阵列含有与核酸样品中靶核酸的子序列互补的寡核苷酸探针。 在一个实施方案中,所述方法包括提供包含一个或多个靶基因的RNA转录物或衍生自RNA转录物的核酸的靶核酸库,将所述核酸库与固定在表面上的寡核苷酸探针阵列杂交,其中 包含超过100种不同寡核苷酸的阵列和每种不同的寡核苷酸被定位在表面的预定区域中,不同寡核苷酸的密度大于每1cm 2大约60个不同的寡核苷酸,并且寡核苷酸探针与RNA转录物互补 衍生自RNA转录物的核酸; 并定量阵列中的杂交核酸。