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1. 发明申请

WO2010140066A2 METHOD OF ALTERING NUCLEIC ACIDS 审中-公开
标题翻译：改变核酸的方法
公开(公告)号：WO2010140066A2
公开(公告)日：2010-12-09
申请号：PCT/IB2010/002396
申请日：2010-06-04
申请人： GENE BRIDGES GMBH , STEWART, Adrian, Francis , IACONE, Roberto , MARESCA, Marcello , ANASTASSIADIS, Konstantinos
发明人： STEWART, Adrian, Francis , IACONE, Roberto , MARESCA, Marcello , ANASTASSIADIS, Konstantinos
CPC分类号： C12N15/102 , C12N15/111 , C12N15/113 , C12N15/90 , C12N2310/124 , C12N2320/50
摘要： The invention provides a method for altering the sequence of a target nucleic acid molecule to incorporate a replacement nucleic acid sequence. The method comprising the steps of: a) introducing a nucleic acid fragment into the target nucleic acid molecule by homologous recombination, wherein the nucleic acid fragment comprises: i) a first region of homology to the target nucleic acid molecule; ii) a 5' splice site; iii) a selectable marker; iv) a 3' splice site; v) a second region of homology to the target nucleic acid molecule; wherein components i) to v) are ordered from 5' to 3'; and the replacement nucleic acid sequence is positioned within or between the first and second regions of homology but external of the splice sites; b) selecting for the introduction of the nucleic acid fragment using the selectable marker; c) incubating the product of step b) under conditions suitable for a splicing reaction to occur such that the selectable marker is excised and from the target nucleic acid, and thereby generating the desired altered target nucleic acid sequence.
摘要翻译：本发明提供了改变靶核酸分子序列以掺入置换核酸序列的方法。该方法包括以下步骤：a）通过同源重组将核酸片段引入靶核酸分子，其中所述核酸片段包含：i）与靶核酸分子同源的第一区域; ii）5'剪接位点; iii）可选标记; iv）3'剪接位点; v）与靶核酸分子同源的第二区域; 其中组分i）至v）从5'到3'被排序; 并且所述置换的核酸序列位于所述第一和第二同源区之内或之间，但位于所述剪接位点的外部; b）使用选择性标记选择引入核酸片段; c）在适合于剪接反应的条件下孵育步骤b）的产物，使得切除选择性标记并从靶核酸中产生所需的改变的靶核酸序列。

2. 发明申请

WO2010113031A3 METHOD OF ALTERING NUCLEIC ACIDS 审中-公开
公开(公告)号：WO2010113031A3
公开(公告)日：2010-10-07
申请号：PCT/IB2010/000893
申请日：2010-03-30
申请人： GENE BRIDGES GMBH , STEWART, Adrian, Francis , ZHANG, Youming , MARESCA, Marcello , KRANZ, Harald , NOLL, Stephan
发明人： STEWART, Adrian, Francis , ZHANG, Youming , MARESCA, Marcello , KRANZ, Harald , NOLL, Stephan
IPC分类号： C12N15/10 , C12N15/90
摘要： The invention provides a method for altering the sequence of a target nucleic acid molecule, said method comprising the steps of: a) introducing a nucleic acid fragment into the target nucleic acid molecule by homologous recombination, wherein the nucleic acid fragment comprises: i) a first region of homology to the target nucleic acid molecule; ii) a first recognition sequence for a Type IIS nuclease; iii) a selectable marker; iv) a second recognition sequence for a Type IIS nuclease; v) a second region of homology to the target nucleic acid molecule; wherein components i) to v) are ordered from 5' to 3'; and a replacement nucleic acid sequence positioned between the first and second regions of homology but flanking the recognition sequences for the Type IIS nucleases; b) selecting for the incorporation of the replacement nucleic acid using the selectable marker, and c) cleaving the product of step b) with Type IIS nucleases such that the selectable marker is excised, to produce a linear target fragment including the desired replacement sequence.

3. 发明申请

WO2002062988A2 RECOMBINATION METHOD 审中-公开
标题翻译：重组方法
公开(公告)号：WO2002062988A2
公开(公告)日：2002-08-15
申请号：PCT/IB2002/001415
申请日：2002-02-08
申请人： GENE BRIDGES GMBH , MUIJRERS, Joep, P., P. , ZHANG, Youming , STEWART, Adrian, Francis
发明人： MUIJRERS, Joep, P., P. , ZHANG, Youming , STEWART, Adrian, Francis
IPC分类号： C12N15/00
CPC分类号： C12N15/102
摘要： The invention relates to a novel method for altering the sequence of a nucleic acid molecule using repair recombination in a simple one component system. The frequency of the recombination reaction is high, allowing a range of feasible selection strategies to identify successful recombination events. The method involves the steps of bringing a first nucleic acid molecule into contact with a second nucleic acid molecule in the presence of a phage annealing protein into contact with a second nucleic acid molecule in the presence of a phage annealing protein, or a functional equivalent or fragment thereof, wherein said first nucleic acid molecule comprises at least two regions of shared sequence homology with the second nucleic acid molecule, under conditions suitable for repair recombination to occur between said first and second nucleic acid molecules; ad selecting a nucleic acid molecule whose sequence has been altered so as to include sequence from said second nucleic acid molecule.
摘要翻译：本发明涉及一种在简单的单一成分系统中使用修复重组改变核酸分子的序列的新方法。重组反应的频率很高，允许一系列可行的选择策略来鉴定成功的重组事件。该方法包括以下步骤：在噬菌体退火蛋白存在下，使第一核酸分子与第二核酸分子接触，以在噬菌体退火蛋白或功能等价物或存在下与第二核酸分子接触，其中所述第一核酸分子在适于在所述第一和第二核酸分子之间发生修复重组的条件下包含与所述第二核酸分子具有共享序列同源性的至少两个区域; 选择其序列已被改变以便包括来自所述第二核酸分子的序列的核酸分子。

4. 发明申请

WO2017174753A1 ADENOVIRAL VECTORS 审中-公开
标题翻译：腺病毒载体
公开(公告)号：WO2017174753A1
公开(公告)日：2017-10-12
申请号：PCT/EP2017/058306
申请日：2017-04-06
申请人： GENE BRIDGES GMBH
发明人： STEWART, Adrian, Francis , FU, Jun , EHRHARDT, Anja , EHRKE-SCHULZ, Eric , ZHANG, Wenli
IPC分类号： C12N15/86 , C07K14/005
摘要： The invention relates to adenoviral vectors, cells for use in generating adenoviral vectors, methods for generating adenoviral vectors, and therapeutic uses of adenoviral vectors in gene therapy, tumour therapy and as vaccines.
摘要翻译：本发明涉及腺病毒载体，用于产生腺病毒载体的细胞，产生腺病毒载体的方法以及腺病毒载体在基因治疗，肿瘤治疗和疫苗中的治疗用途。

5. 发明申请

WO2009104094A2 METHOD OF NUCLEIC ACID RECOMBINATION 审中-公开
标题翻译：核酸重组方法
公开(公告)号：WO2009104094A2
公开(公告)日：2009-08-27
申请号：PCT/IB2009/000488
申请日：2009-02-20
申请人： GENE BRIDGES GMBH , MARESCA, Marcello , ERLER, Axel, Steffen , FU, Jun , SEIBERT, Philipp, Martin , STEWART, Adrian, Francis , ZHANG, Youming
发明人： MARESCA, Marcello , ERLER, Axel, Steffen , FU, Jun , SEIBERT, Philipp, Martin , STEWART, Adrian, Francis , ZHANG, Youming
IPC分类号： C12N15/09
CPC分类号： C12N15/1082
摘要： The invention provides a method for inserting a single stranded replacement nucleic acid into a target nucleic acid, said method comprising the steps of: a) generating a single stranded replacement nucleic acid from a double stranded nucleic acid, wherein the double stranded nucleic acid is adapted at one or both of its 5' ends such that preferential degradation of one strand and/or strand separation generates the single stranded replacement nucleic acid, wherein the single stranded replacement nucleic acid comprises a 5' region that is identical to sequence on the target nucleic acid, a 3' region that is identical to sequence on the target nucleic acid and optionally a replacement region between the 5' and 3' regions that is not identical to sequence on the target nucleic acid, b) exposing the target nucleic acid to the single stranded replacement nucleic acid under conditions suitable for recombination to occur between the single stranded replacement nucleic acid and the target nucleic acid, and c) selecting a target nucleic acid whose sequence has been altered by inclusion of said single stranded replacement nucleic acid. Other methods for modifying target nucleic acids are also provided.
摘要翻译：本发明提供了将单链置换核酸插入靶核酸的方法，所述方法包括以下步骤：a）从双链核酸产生单链置换核酸，其中双链核酸适应在其5'末端的一个或两个上，使得一条链和/或链分离的优先降解产生单链置换核酸，其中单链置换核酸包含与靶核酸上的序列相同的5'区酸，3'区，其与靶核酸上的序列相同，并且任选地在与靶核酸上的序列不相同的5'和3'区之间的置换区，b）将靶核酸暴露于单链置换核酸在适于在单链置换核酸和靶核之间发生重组的条件下进行并且c）选择通过包含所述单链置换核酸改变其序列的靶核酸。还提供了用于修饰靶核酸的其它方法。

6. 发明申请

WO2002062988A3 RECOMBINATION METHOD 审中-公开
公开(公告)号：WO2002062988A3
公开(公告)日：2002-08-15
申请号：PCT/IB2002/001415
申请日：2002-02-08
申请人： GENE BRIDGES GMBH , MUIJRERS, Joep, P., P. , ZHANG, Youming , STEWART, Adrian, Francis
发明人： MUIJRERS, Joep, P., P. , ZHANG, Youming , STEWART, Adrian, Francis
IPC分类号： C12N15/00
摘要： The invention relates to a novel method for altering the sequence of a nucleic acid molecule using repair recombination in a simple one component system. The frequency of the recombination reaction is high, allowing a range of feasible selection strategies to identify successful recombination events. The method involves the steps of bringing a first nucleic acid molecule into contact with a second nucleic acid molecule in the presence of a phage annealing protein into contact with a second nucleic acid molecule in the presence of a phage annealing protein, or a functional equivalent or fragment thereof, wherein said first nucleic acid molecule comprises at least two regions of shared sequence homology with the second nucleic acid molecule, under conditions suitable for repair recombination to occur between said first and second nucleic acid molecules; ad selecting a nucleic acid molecule whose sequence has been altered so as to include sequence from said second nucleic acid molecule.

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