公开(公告)号：US09777290B2

公开(公告)日：2017-10-03

申请号：US14426329

申请日：2013-09-23

申请人： Lonza Biologics PLC.

发明人： Tom Payne ,  Robert Young ,  Marc Feary

IPC分类号： C12N15/85

CPC分类号： C12N15/85 ,  C12N2710/16122 ,  C12N2830/15 ,  C12N2830/60

摘要： The present invention relates to expression vectors for the heterologous expression of a nucleic acid sequence of interest in mammalian cells, the vectors comprising a chimeric promoter regulatory sequence being operably linked to a nucleic acid sequence to be expressed, wherein the chimeric promoter regulatory sequence comprises a cytomegalovirus promoter sequence derived from murine cytomegalovirus or from human cytomegalovirus and being operably linked to the transcriptional start site of the nucleic acid sequence to be expressed; and a cytomegalovirus upstream region and/or enhancer sequence derived from human and/or the simian cytomegalovirus, wherein the upstream region and/or enhancer sequence is located 5′ of and operably linked to the murine or the human promoter sequence, and wherein the chimeric promoter regulatory sequence comprises sequence elements being derived from at least two of the group consisting of murine cytomegalovirus, human cytomegalovirus and simian cytomegalovirus. In particular embodiments, the chimeric promoter regulatory sequence comprises sequence elements derived from the murine or the human cytomegalovirus IE1 promoter and from the human and/or the simian cytomegalovirus IE1 region. The invention also relates to mammalian host cells transfected with such expression vectors, a method for heterologous expression of a nucleic acid sequence in a mammalian host cell by employing such expression vectors, and the use of such expression vectors for the heterologous expression of a nucleic acid sequence.

公开(公告)号：US10294491B2

公开(公告)日：2019-05-21

申请号：US15687835

申请日：2017-08-28

申请人： Lonza Biologics PLC.

发明人： Tom Payne ,  Robert Young ,  Marc Feary

IPC分类号： C12N15/85

摘要： The present invention relates to expression vectors for the heterologous expression of a nucleic acid sequence of interest in mammalian cells, the vectors comprising a chimeric promoter regulatory sequence being operably linked to a nucleic acid sequence to be expressed, wherein the chimeric promoter regulatory sequence comprises a cytomegalovirus promoter sequence derived from murine cytomegalovirus or from human cytomegalovirus and being operably linked to the transcriptional start site of the nucleic acid sequence to be expressed; and a cytomegalovirus upstream region and/or enhancer sequence derived from human and/or the simian cytomegalovirus, wherein the upstream region and/or enhancer sequence is located 5′ of and operably linked to the murine or the human promoter sequence, and wherein the chimeric promoter regulatory sequence comprises sequence elements being derived from at least two of the group consisting of murine cytomegalovirus, human cytomegalovirus and simian cytomegalovirus. In particular embodiments, the chimeric promoter regulatory sequence comprises sequence elements derived from the murine or the human cytomegalovirus IE1 promoter and from the human and/or the simian cytomegalovirus IE1 region. The invention also relates to mammalian host cells transfected with such expression vectors, a method for heterologous expression of a nucleic acid sequence in a mammalian host cell by employing such expression vectors, and the use of such expression vectors for the heterologous expression of a nucleic acid sequence.

公开(公告)号：US20150218584A1

公开(公告)日：2015-08-06

申请号：US14426329

申请日：2013-09-23

申请人： Lonza Biologics PLC.

发明人： Tom Payne ,  Robert Young ,  Marc Feary

IPC分类号： C12N15/85

CPC分类号： C12N15/85 ,  C12N2710/16122 ,  C12N2830/15 ,  C12N2830/60

摘要： The present invention relates to expression vectors for the heterologous expression of a nucleic acid sequence of interest in mammalian cells, the vectors comprising a chimeric promoter regulatory sequence being operably linked to a nucleic acid sequence to be expressed, wherein the chimeric promoter regulatory sequence comprises a cytomegalovirus promoter sequence derived from murine cytomegalovirus or from human cytomegalovirus and being operably linked to the transcriptional start site of the nucleic acid sequence to be expressed; and a cytomegalovirus upstream region and/or enhancer sequence derived from human and/or the simian cytomegalovirus, wherein the upstream region and/or enhancer sequence is located 5′ of and operably linked to the murine or the human promoter sequence, and wherein the chimeric promoter regulatory sequence comprises sequence elements being derived from at least two of the group consisting of murine cytomegalovirus, human cytomegalovirus and simian cytomegalovirus. In particular embodiments, the chimeric promoter regulatory sequence comprises sequence elements derived from the murine or the human cytomegalovirus IE1 promoter and from the human and/or the simian cytomegalovirus IE1 region. The invention also relates to mammalian host cells transfected with such expression vectors, a method for heterologous expression of a nucleic acid sequence in a mammalian host cell by employing such expression vectors, and the use of such expression vectors for the heterologous expression of a nucleic acid sequence.

摘要翻译： 本发明涉及用于在哺乳动物细胞中异源表达目的核酸序列的表达载体，所述载体包含可操作地连接于待表达的核酸序列的嵌合启动子调节序列，其中所述嵌合启动子调控序列包含 源自鼠巨细胞病毒或来自人巨细胞病毒的巨细胞病毒启动子序列，并且与要表达的核酸序列的转录起始位点可操作地连接; 和源自人和/或猿巨细胞病毒的巨细胞病毒上游区和/或增强子序列，其中所述上游区和/或增强子序列位于鼠或人启动子序列的5'并且可操作地连接到鼠和人启动子序列，并且其中嵌合 启动子调节序列包含源自由鼠巨细胞病毒，人巨细胞病毒和猿巨细胞病毒组成的组中的至少两个的序列元件。 在具体实施方案中，嵌合启动子调节序列包含衍生自鼠或人巨细胞病毒IE1启动子和来自人和/或猿巨细胞病毒IE1区的序列元件。 本发明还涉及用这种表达载体转染的哺乳动物宿主细胞，通过使用这种表达载体在哺乳动物宿主细胞中异源表达核酸序列的方法，以及使用这种表达载体来异源表达核酸 序列。

公开(公告)号：US10669541B2

公开(公告)日：2020-06-02

申请号：US14910267

申请日：2014-07-30

申请人： LONZA BIOLOGICS PLC

发明人： Marc Feary ,  James Rance ,  Robert Young ,  Elizabeth C. Sayer ,  Christopher M. Smales

IPC分类号： C12N15/113 ,  C12P21/00 ,  C12N15/11 ,  C07K16/00

摘要： Methods for generating mammalian cells characterized by an increased concentration of at least one of miR-15, miR-16, and miR-34 for producing proteins at an industrial scale are provided. Methods for using the mammalian cells, in particular for the production of proteins of interest, are also provided.

公开(公告)号：US20200002721A1

公开(公告)日：2020-01-02

申请号：US16372738

申请日：2019-04-02

申请人： Lonza Biologics PLC

发明人： Tom Payne ,  Robert Young ,  Marc Feary

IPC分类号： C12N15/85

摘要： The present invention relates to expression vectors for the heterologous expression of a nucleic acid sequence of interest in mammalian cells, the vectors comprising a chimeric promoter regulatory sequence being operably linked to a nucleic acid sequence to be expressed, wherein the chimeric promoter regulatory sequence comprises a cytomegalovirus promoter sequence derived from murine cytomegalovirus or from human cytomegalovirus and being operably linked to the transcriptional start site of the nucleic acid sequence to be expressed; and a cytomegalovirus upstream region and/or enhancer sequence derived from human and/or the simian cytomegalovirus, wherein the upstream region and/or enhancer sequence is located 5′ of and operably linked to the murine or the human promoter sequence, and wherein the chimeric promoter regulatory sequence comprises sequence elements being derived from at least two of the group consisting of murine cytomegalovirus, human cytomegalovirus and simian cytomegalovirus. In particular embodiments, the chimeric promoter regulatory sequence comprises sequence elements derived from the murine or the human cytomegalovirus IE1 promoter and from the human and/or the simian cytomegalovirus IE1 region. The invention also relates to mammalian host cells transfected with such expression vectors, a method for heterologous expression of a nucleic acid sequence in a mammalian host cell by employing such expression vectors, and the use of such expression vectors for the heterologous expression of a nucleic acid sequence.

公开(公告)号：US20170356007A1

公开(公告)日：2017-12-14

申请号：US15687835

申请日：2017-08-28

申请人： Lonza Biologics PLC.

发明人： Tom Payne ,  Robert Young ,  Marc Feary

IPC分类号： C12N15/85

CPC分类号： C12N15/85 ,  C12N2710/16122 ,  C12N2830/15 ,  C12N2830/60

摘要： The present invention relates to expression vectors for the heterologous expression of a nucleic acid sequence of interest in mammalian cells, the vectors comprising a chimeric promoter regulatory sequence being operably linked to a nucleic acid sequence to be expressed, wherein the chimeric promoter regulatory sequence comprises a cytomegalovirus promoter sequence derived from murine cytomegalovirus or from human cytomegalovirus and being operably linked to the transcriptional start site of the nucleic acid sequence to be expressed; and a cytomegalovirus upstream region and/or enhancer sequence derived from human and/or the simian cytomegalovirus, wherein the upstream region and/or enhancer sequence is located 5′ of and operably linked to the murine or the human promoter sequence, and wherein the chimeric promoter regulatory sequence comprises sequence elements being derived from at least two of the group consisting of murine cytomegalovirus, human cytomegalovirus and simian cytomegalovirus. In particular embodiments, the chimeric promoter regulatory sequence comprises sequence elements derived from the murine or the human cytomegalovirus IE1 promoter and from the human and/or the simian cytomegalovirus IE1 region. The invention also relates to mammalian host cells transfected with such expression vectors, a method for heterologous expression of a nucleic acid sequence in a mammalian host cell by employing such expression vectors, and the use of such expression vectors for the heterologous expression of a nucleic acid sequence.