公开(公告)号：WO2016033116A1

公开(公告)日：2016-03-03

申请号：PCT/US2015/046811

申请日：2015-08-25

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： ROGERS, Jeffrey ,  LAM, Bianca ,  GUO, Joanna ,  LI, Lushen

IPC: C12N9/99 ,  C12N9/12

CPC classification number: C12N9/1276 ,  C12N9/12 ,  C12N9/99 ,  C12P19/34 ,  C12Y207/07049

Abstract: The present invention provides reversibly inactivated reverse transcriptase enzymes, particularly those having increased thermostability and/or thermoreactivity and compositions, methods and kits that include such enzymes, for the reverse transcription of nucleic acid molecules. Also provided are compositions and methods for the reactivation of reversibly inactivated reverse transcriptases. More particularly, the present invention relates to compositions and methods that can increase the speed of reactivation and stabilize reactivation of chemically modified reverse transcriptase enzymes prior to, or as part of a reverse transcription reaction. As compared to existing compositions and methods, the reversibly inactivated reverse transcriptase enzymes of the present invention provide for a significant reduction in non-specific reverse transcription from template nucleic acid molecules.

Abstract translation: 本发明提供了可逆地灭活的逆转录酶，特别是那些具有增加的热稳定性和/或热反应性的组合物，包含这些酶的组合物，方法和试剂盒，用于逆转录核酸分子。 还提供了用于重新激活可逆失活的逆转录酶的组合物和方法。 更具体地说，本发明涉及可以在逆转录反应之前或作为逆转录反应的一部分期间增加再活化速度并稳定化学修饰的逆转录酶的再活化的组合物和方法。 与现有的组合物和方法相比，本发明的可逆失活的逆转录酶提供了模板核酸分子的非特异性逆转录的显着降低。

公开(公告)号：WO2013166339A1

公开(公告)日：2013-11-07

申请号：PCT/US2013/039351

申请日：2013-05-02

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： VASU, Sanjay, K. ,  CHIOU, Henry, C. ,  ROGERS, Jeffrey ,  CISNEROS, Maria ,  LI, Jingqiu ,  LIU, Chao, Yan ,  JONES, Meredith

IPC: C12N5/00

CPC classification number: C07K14/505 ,  C12N5/005 ,  C12N2500/30 ,  C12N2501/065 ,  C12N2501/999 ,  C12N2510/02 ,  C12N2511/00

Abstract: The present invention is directed generally to cell culture media (particularly serum free, non animal derived, and/or chemically defined media) which are useful for introducing macromolecules and compounds (e.g., nucleic acid molecules) into cells (e.g., eukaryotic cells). According to the invention, such introduction can take place in the presence of said medium. Cells containing such introduced materials can then be cultured in the medium and the effect of the introduced materials on the cells can be measured or determined. In particular, the invention allows introduction of nucleic acid molecules (e.g., vectors) into cells (particularly eukaryotic cells) and expression of proteins encoded by the nucleic acid molecules in the cells. The invention obviates the need to change the cell culture medium each time a different procedure is performed with the cells (e.g., culturing cells vs. transfecting cells). The invention also relates to compositions and kits useful for culturing and transforming/transfecting cells.

Abstract translation: 本发明一般涉及可用于将大分子和化合物（例如，核酸分子）引入细胞（例如真核细胞）中的细胞培养基（特别是无血清，非动物衍生的和/或化学上限定的培养基）。 根据本发明，这种引入可以在所述介质的存在下进行。 然后可以在培养基中培养含有这种引入的物质的细胞，并且可以测量或测定引入的细胞对细胞的影响。 特别地，本发明允许将核酸分子（例如，载体）引入细胞（特别是真核细胞）中，以及由细胞中核酸分子编码的蛋白质的表达。 本发明避免了每当与细胞（例如培养细胞与转染细胞）进行不同程序时改变细胞培养基的需要。 本发明还涉及可用于培养和转化/转染细胞的组合物和试剂盒。

公开(公告)号：WO2015112767A2

公开(公告)日：2015-07-30

申请号：PCT/US2015/012534

申请日：2015-01-22

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： ROGERS, Jeffrey ,  POTTER, Jason

IPC: C12N9/22

CPC classification number: C12N9/1276 ,  C12N15/1096 ,  C12P19/34 ,  C12Y207/07049

Abstract: The invention provides novel reverse transcriptases (RTs) with desirable properties such as increased thermostability, increased thermoreactivity and/or increased resistance to inhibitors. In certain embodiments, the invention provides methods of producing, amplifying and/or sequencing nucleic acid molecules (particularly cDNA molecules) using kits, compositions and/or reactions mixtures containing such novel reverse transcriptase enzymes.

Abstract translation: 本发明提供具有所需性质的新型逆转录酶（RT），例如增加的热稳定性，增加的热反应性和/或增加的对抑制剂的抗性。 在某些实施方案中，本发明提供使用包含这种新型逆转录酶的试剂盒，组合物和/或反应混合物来产生，扩增和/或测序核酸分子（特别是cDNA分子）的方法。

公开(公告)号：WO2014066404A2

公开(公告)日：2014-05-01

申请号：PCT/US2013/066204

申请日：2013-10-22

Applicant: LIFE TECHNOLOGIES CORPORATION

Inventor： ROGERS, Jeffrey ,  FLETCHER, Julia ,  LOWITZ, Kevin

IPC: C12N15/09

CPC classification number: C12N15/67 ,  C07K2/00 ,  C07K2319/40 ,  C07K2319/60 ,  C12N9/2471 ,  C12N15/1086 ,  C12N15/63 ,  C12N15/70 ,  C12P21/00

Abstract: The present disclosure describes a novel recombinant gene expression system and a method of its use. The recombinant gene expression system is adapted to enable a user to monitor the successful expression of a protein in real time by allowing a small portion (e.g., less than 10%) of the target protein to be expressed as a fusion with a readily detectable reporter. The recombinant gene expression system further allows a user to select a clone that exhibits high, medium or low expression by providing a random ribosomal binding site having the nucleotide sequence N 4 R 6 N X , where N is A, T, G, or C, where R is A or G, and where x is an integer from 6 to 11 upstream of the cloned gene. By selecting a clone exhibiting a desired relative level of reporter gene expression, a user can tailor the expression level of the desired protein.

Abstract translation: 本公开描述了一种新的重组基因表达系统及其使用方法。 重组基因表达系统适于使用户能够通过使目标蛋白质的一小部分（例如，小于10％）表达为与容易检测的报告基团的融合物来实时监测蛋白质的成功表达 。 重组基因表达系统还允许用户通过提供具有核苷酸序列N4R6NX的随机核糖体结合位点来选择表现出高，中或低表达的克隆，其中N是A，T，G或C，其中R是A 或G，并且其中x是克隆基因上游6至11的整数。 通过选择表现出所需相对水平的报告基因表达的克隆，用户可以调整所需蛋白质的表达水平。

公开(公告)号：EP2844737A1

公开(公告)日：2015-03-11

申请号：EP13722946.4

申请日：2013-05-02

Applicant: Life Technologies Corporation

Inventor： VASU, Sanjay, K. ,  CHIOU, Henry, C. ,  ROGERS, Jeffrey ,  CISNEROS, Maria ,  LI, Jingqiu ,  LIU, Chao, Yan ,  JONES, Meredith

IPC: C12N5/00

CPC classification number: C07K14/505 ,  C12N5/005 ,  C12N2500/30 ,  C12N2501/065 ,  C12N2501/999 ,  C12N2510/02 ,  C12N2511/00

Abstract: The present invention is directed generally to cell culture media (particularly serum free, non animal derived, and/or chemically defined media) which are useful for introducing macromolecules and compounds (e.g., nucleic acid molecules) into cells (e.g., eukaryotic cells). According to the invention, such introduction can take place in the presence of said medium. Cells containing such introduced materials can then be cultured in the medium and the effect of the introduced materials on the cells can be measured or determined. In particular, the invention allows introduction of nucleic acid molecules (e.g., vectors) into cells (particularly eukaryotic cells) and expression of proteins encoded by the nucleic acid molecules in the cells. The invention obviates the need to change the cell culture medium each time a different procedure is performed with the cells (e.g., culturing cells vs. transfecting cells). The invention also relates to compositions and kits useful for culturing and transforming/transfecting cells.

公开(公告)号：EP4148122A1

公开(公告)日：2023-03-15

申请号：EP22186964.7

申请日：2013-05-02

Applicant: Life Technologies Corporation

Inventor： VASU, Sanjay, K. ,  CHIOU, Henry, C. ,  ROGERS, Jeffrey ,  CISNEROS, Maria ,  LI, Jingqiu ,  LIU, Chao, Yan ,  JONES, Meredith

IPC: C12N5/00 ,  C12N5/0793

Abstract: The present invention is directed generally to cell culture media (particularly serum free, non animal derived, and/or chemically defined media) which are useful for introducing macromolecules and compounds (e.g., nucleic acid molecules) into cells (e.g., eukaryotic cells). According to the invention, such introduction can take place in the presence of said medium. Cells containing such introduced materials can then be cultured in the medium and the effect of the introduced materials on the cells can be measured or determined. In particular, the invention allows introduction of nucleic acid molecules (e.g., vectors) into cells (particularly eukaryotic cells) and expression of proteins encoded by the nucleic acid molecules in the cells. The invention obviates the need to change the cell culture medium each time a different procedure is performed with the cells (e.g., culturing cells vs. transfecting cells). The invention also relates to compositions and kits useful for culturing and transforming/transfecting cells.

公开(公告)号：EP3404095A1

公开(公告)日：2018-11-21

申请号：EP18162843.9

申请日：2013-05-02

Applicant: Life Technologies Corporation

Inventor： VASU, Sanjay, K. ,  CHIOU, Henry, C. ,  ROGERS, Jeffrey ,  CISNEROS, Maria ,  LI, Jingqiu ,  LIU, Chao, Yan ,  JONES, Meredith

IPC: C12N5/00

CPC classification number: C07K14/505 ,  C12N5/005 ,  C12N5/0619 ,  C12N2500/30 ,  C12N2501/065 ,  C12N2501/999 ,  C12N2510/02 ,  C12N2511/00

Abstract: The present invention is directed generally to cell culture media (particularly serum free, non animal derived, and/or chemically defined media) which are useful for introducing macromolecules and compounds (e.g., nucleic acid molecules) into cells (e.g., eukaryotic cells). According to the invention, such introduction can take place in the presence of said medium. Cells containing such introduced materials can then be cultured in the medium and the effect of the introduced materials on the cells can be measured or determined. In particular, the invention allows introduction of nucleic acid molecules (e.g., vectors) into cells (particularly eukaryotic cells) and expression of proteins encoded by the nucleic acid molecules in the cells. The invention obviates the need to change the cell culture medium each time a different procedure is performed with the cells (e.g., culturing cells vs. transfecting cells). The invention also relates to compositions and kits useful for culturing and transforming/transfecting cells.

公开(公告)号：EP2844737B1

公开(公告)日：2018-03-21

申请号：EP13722946.4

申请日：2013-05-02

Applicant: Life Technologies Corporation

Inventor： VASU, Sanjay, K. ,  CHIOU, Henry, C. ,  ROGERS, Jeffrey ,  CISNEROS, Maria ,  LI, Jingqiu ,  LIU, Chao, Yan ,  JONES, Meredith

IPC: C12N5/071 ,  C12N5/00 ,  C07K14/505

CPC classification number: C07K14/505 ,  C12N5/005 ,  C12N5/0619 ,  C12N2500/30 ,  C12N2501/065 ,  C12N2501/999 ,  C12N2510/02 ,  C12N2511/00

Abstract: High-Yield mammalian transient expression systems can include a cell culture media (particularly serum free, non animal derived, and/or chemically defined media) for introducing macromolecules and compounds (e.g., nucleic acid molecules) into cells (e.g., eukaryotic cells). Cells containing such introduced materials can then be cultured in the cell culture media. In particular, the invention allows introduction of nucleic acid molecules (e.g., vectors) into cells (particularly mammalian cells) and expression of proteins encoded by the nucleic acid molecules in the cells. The invention obviates the need to change the cell culture medium each time a different procedure is performed with the cells (e.g., culturing cells vs. transfecting cells). The invention also relates to compositions and kits useful for culturing and transforming/transfecting cells.

公开(公告)号：EP3097190A2

公开(公告)日：2016-11-30

申请号：EP15702365.6

申请日：2015-01-22

Applicant: Life Technologies Corporation

Inventor： ROGERS, Jeffrey ,  POTTER, Jason

IPC: C12N9/22 ,  C12N9/12

CPC classification number: C12N9/1276 ,  C12N15/1096 ,  C12P19/34 ,  C12Y207/07049

Abstract: The invention provides novel reverse transcriptases (RTs) with desirable properties such as increased thermostability, increased thermoreactivity and/or increased resistance to inhibitors. In certain embodiments, the invention provides methods of producing, amplifying and/or sequencing nucleic acid molecules (particularly cDNA molecules) using kits, compositions and/or reactions mixtures containing such novel reverse transcriptase enzymes.

公开(公告)号：EP3186374A1

公开(公告)日：2017-07-05

申请号：EP15757431.0

申请日：2015-08-25

Applicant: Life Technologies Corporation

Inventor： ROGERS, Jeffrey ,  LAM, Bianca ,  GUO, Joanna ,  LI, Lushen

IPC: C12N9/99 ,  C12N9/12

CPC classification number: C12N9/1276 ,  C12N9/12 ,  C12N9/99 ,  C12P19/34 ,  C12Y207/07049

Abstract: The present invention provides reversibly inactivated reverse transcriptase enzymes, particularly those having increased thermostability and/or thermoreactivity and compositions, methods and kits that include such enzymes, for the reverse transcription of nucleic acid molecules. Also provided are compositions and methods for the reactivation of reversibly inactivated reverse transcriptases. More particularly, the present invention relates to compositions and methods that can increase the speed of reactivation and stabilize reactivation of chemically modified reverse transcriptase enzymes prior to, or as part of a reverse transcription reaction. As compared to existing compositions and methods, the reversibly inactivated reverse transcriptase enzymes of the present invention provide for a significant reduction in non-specific reverse transcription from template nucleic acid molecules.

Abstract translation: 本发明提供可逆性失活的逆转录酶，特别是具有增加的热稳定性和/或热反应性的那些酶，以及包含这种酶的组合物，方法和试剂盒，用于逆转录核酸分子。 还提供了用于可逆性失活的逆转录酶的再激活的组合物和方法。 更具体地说，本发明涉及在逆转录反应之前或作为逆转录反应的一部分可增加再激活速度并稳定化学修饰的逆转录酶的再激活的组合物和方法。 与现有的组合物和方法相比，本发明的可逆性失活的逆转录酶使来自模板核酸分子的非特异性逆转录显着减少。