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1. 发明申请

WO2013135729A1 RECOMBINATION SYSTEM 审中-公开
标题翻译：重塑系统
公开(公告)号：WO2013135729A1
公开(公告)日：2013-09-19
申请号：PCT/EP2013/055048
申请日：2013-03-12
申请人： DSM IP ASSETS B.V.
发明人： PEIJ, VAN, Noël Nicolaas Maria Elisabeth , BEISHUIZEN, Martina , ARENDSEN, Yvonne Johannes Odilia
IPC分类号： C12N15/63 , C12N15/90 , C12N15/00 , C12N5/00
CPC分类号： C12N15/85 , C12N15/80 , C12N15/81 , C12N15/82 , C12N15/902
摘要： The present invention relates to a method for carrying out recombination at a target locus, which method comprises: providing two or more nucleic acids which, when taken together, comprise: (a) sequences capable of homologous recombination with sequences flanking the target locus; (b) two or more site-specific recombination sites; (c) a sequence encoding a recombinase which recognizes the site-specific recombination sites; and (d) a sequence encoding a marker, wherein the two or more nucleic acids are capable of homologous recombination with each other so as to give rise to a single nucleic acid, and wherein at least two of the two or more nucleic acids each comprise a sequence encoding a non-functional portion of the marker; and recombining the said two or more nucleic acids with each other and with the sequences flanking the target locus so that a contiguous nucleic acid sequence encoding a functional marker and the sequence encoding the recombinase are inserted at the target locus, said marker-encoding and/or recombinase- encoding sequence being flanked by at least two site-specific recombination sites and the said site-specific recombination sites being flanked by the sequences capable of homologous recombination with sequences flanking the target locus.
摘要翻译：本发明涉及在目标基因座上进行重组的方法，该方法包括：提供两种或多种核酸，当它们一起时，它们包含：（a）能够与靶基因座侧翼的序列同源重组的序列; （b）两个或多个位点特异性重组位点; （c）编码识别位点特异性重组位点的重组酶的序列; 和（d）编码标记的序列，其中所述两个或更多个核酸能够彼此进行同源重组，以产生单个核酸，并且其中所述两个或更多个核酸中的至少两个各自包含编码标记的非功能部分的序列; 并且将所述两种或多种核酸彼此重组，并与靶基因座侧翼的序列重组，使得编码功能性标记的连续核酸序列和编码重组酶的序列插入靶标位点，所述标记编码和/ 或重组酶编码序列，其侧翼为至少两个位点特异性重组位点，并且所述位点特异性重组位点侧翼为能够与靶基因座侧翼序列同源重组的序列。

2. 发明申请

WO2013135732A1 RASAMSONIA TRANSFORMANTS 审中-公开
标题翻译： RASAMSONIA变种
公开(公告)号：WO2013135732A1
公开(公告)日：2013-09-19
申请号：PCT/EP2013/055051
申请日：2013-03-12
申请人： DSM IP ASSETS B.V.
发明人： LOS, Alrik Pieter , ARENDSEN, Yvonne Johannes Odilia , VOLLEBREGT, Adrianus Wilhelmus Hermanus
IPC分类号： C12N15/90 , C12N5/00 , C07K14/00
CPC分类号： C12N15/80 , C12N9/2437 , C12N9/58 , C12N15/905 , C12N2800/30 , C12Y302/01004 , C12Y304/23
摘要： A method for carrying out recombination at a target locus in a Rasamsonia cell, which method comprises: - providing two or more nucleic acids which, when taken together, comprise: (a) sequences capable of homologous recombination with sequences flanking the target locus; (b) two or more site-specific recombination sites; (c) a sequence encoding a recombinase which recognizes the site-specific recombination sites; and (d) a sequence encoding a marker, wherein the two or more nucleic acids are capable of homologous recombination with each other so as to give rise to a single nucleic acid, and wherein at least two of the two or more nucleic acids each comprise a sequence encoding a non- functional portion of the marker; and - recombining in a Rasamsonia cell the said two or more nucleic acids with each other and with the sequences flanking the target locus so that a contiguous nucleic acid sequence encoding a functional marker and the sequence encoding the recombinase are inserted at the target locus, said marker-encoding and/or recombinase-encoding sequence being flanked by at least two site-specific recombination sites and the said site-specific recombination sites being flanked by the sequences capable of homologous recombination with sequences flanking the target locus, thereby to carry out recombination at a target locus in a Rasamsonia cell. The method may be used to generate marker free Rasamsonia cells.
摘要翻译：该方法包括： - 提供两种或多种核酸，当它们一起时，它们包含：（a）能够与靶基因座侧翼的序列同源重组的序列;一种用于在Rasamsonia细胞中靶标位点进行重组的方法，（b）两个或多个位点特异性重组位点; （c）编码识别位点特异性重组位点的重组酶的序列; 和（d）编码标记的序列，其中所述两个或更多个核酸能够彼此进行同源重组，以产生单个核酸，并且其中所述两个或更多个核酸中的至少两个各自包含编码标记的非功能部分的序列; 以及 - 在所述两个或多个核酸中的Rasamsonia细胞中彼此重叠，并且与靶基因座侧翼的序列重组，使得编码功能性标记的连续核酸序列和编码重组酶的序列插入靶位点，所述标记编码和/或重组酶编码序列侧接至少两个位点特异性重组位点，并且所述位点特异性重组位点位于与靶基因座侧翼序列同源重组的序列的两侧，从而进行重组在Rasamsonia细胞的靶标位点。该方法可用于产生无标记的Rasamsonia细胞。

3. 发明申请

WO2013135728A1 RECOMBINATION SYSTEM 审中-公开
标题翻译：重塑系统
公开(公告)号：WO2013135728A1
公开(公告)日：2013-09-19
申请号：PCT/EP2013/055047
申请日：2013-03-12
申请人： DSM IP ASSETS B.V.
发明人： WIESSENHAAN, Nathalie , KOLEN, Catharina Petronella Antonia Maria , MEIJRINK, Bernard , BOER, Viktor , ROUBOS, Johannes, Andries , ARENDSEN, Yvonne Johannes Odilia
IPC分类号： C12N15/63 , C12N15/90 , C12N15/00 , C12N5/00
CPC分类号： C12N15/102 , C12N15/902 , C12N2800/30
摘要： The present invention relates to a method for carrying out recombination at a target locus, which method comprises: providing two or more nucleic acids which, when taken together, comprise: (a) sequences capable of homologous recombination with sequences flanking the target locus; (b) two or more site-specific recombination sites; and (c) a sequence encoding a recombinase which recognizes the site-specific recombination sites, wherein the two or more nucleic acids are capable of homologous recombination with each other so as to give rise to a single nucleic acid, and wherein at least two of the two or more nucleic acids each comprise a sequence encoding a non-functional portion of the recombinase; and recombining the said two or more nucleic acids with each other and with the sequences flanking the target locus so that a contiguous nucleic acid sequence encoding a functional recombinase is inserted at the target locus, said recombinase-encoding sequence being flanked by at least two site-specific recombination sites and the said site-specific recombination sites being flanked by the sequences capable of homologous recombination with sequences flanking the target locus.
摘要翻译：本发明涉及在目标基因座上进行重组的方法，该方法包括：提供两种或多种核酸，当它们一起时，它们包含：（a）能够与靶基因座侧翼的序列同源重组的序列; （b）两个或多个位点特异性重组位点; 和（c）编码识别位点特异性重组位点的重组酶的序列，其中所述两个或更多个核酸能够彼此进行同源重组以产生单个核酸，并且其中至少两个两个或多个核酸各自包含编码重组酶的非功能部分的序列; 并将所述两种或多种核酸彼此重组，并与靶基因座侧翼的序列重组，使得编码功能重组酶的连续核酸序列插入靶基因座，所述重组酶编码序列侧接至少两个位点特异性重组位点和所述位点特异性重组位点的侧翼是能够与靶基因座侧翼的序列同源重组的序列。

4. 发明授权

EP2825650B1 RECOMBINATION SYSTEM 有权
公开(公告)号：EP2825650B1
公开(公告)日：2019-02-13
申请号：EP13708506.4
申请日：2013-03-12
申请人： DSM IP Assets B.V.
发明人： WIESSENHAAN, Nathalie , KOLEN, Catharina Petronella Antonia Maria , MEIJRINK, Bernard , BOER, Viktor , ROUBOS, Johannes, Andries , ARENDSEN, Yvonne Johannes Odilia
IPC分类号： C12N15/63 , C12N15/90 , C12N15/00 , C12N5/00

5. 发明公开

EP2825659A1 RASAMSONIA TRANSFORMANTS 无效
标题翻译： RASAMSONIA-TRANSFORMANTEN
公开(公告)号：EP2825659A1
公开(公告)日：2015-01-21
申请号：EP13708834.0
申请日：2013-03-12
申请人： DSM IP Assets B.V.
发明人： LOS, Alrik Pieter , ARENDSEN, Yvonne Johannes Odilia , VOLLEBREGT, Adrianus Wilhelmus Hermanus
IPC分类号： C12N15/90 , C12N5/00 , C07K14/00
CPC分类号： C12N15/80 , C12N9/2437 , C12N9/58 , C12N15/905 , C12N2800/30 , C12Y302/01004 , C12Y304/23
摘要： A method for carrying out recombination at a target locus in a
Rasamsonia cell, which method comprises: - providing two or more nucleic acids which, when taken together, comprise: (a) sequences capable of homologous recombination with sequences flanking the target locus; (b) two or more site-specific recombination sites; (c) a sequence encoding a recombinase which recognizes the site-specific recombination sites; and (d) a sequence encoding a marker, wherein the two or more nucleic acids are capable of homologous recombination with each other so as to give rise to a single nucleic acid, and wherein at least two of the two or more nucleic acids each comprise a sequence encoding a non- functional portion of the marker; and - recombining in a
Rasamsonia cell the said two or more nucleic acids with each other and with the sequences flanking the target locus so that a contiguous nucleic acid sequence encoding a functional marker and the sequence encoding the recombinase are inserted at the target locus, said marker-encoding and/or recombinase-encoding sequence being flanked by at least two site-specific recombination sites and the said site-specific recombination sites being flanked by the sequences capable of homologous recombination with sequences flanking the target locus, thereby to carry out recombination at a target locus in a
Rasamsonia cell. The method may be used to generate marker free
Rasamsonia cells.
摘要翻译：一种在Rasamsonia细胞中的目标基因座进行重组的方法，所述方法包括：提供两种或多种核酸，当它们一起包含：（a）能够与靶基因座侧翼的序列同源重组的序列; （b）两个或多个位点特异性重组位点; （c）编码识别位点特异性重组位点的重组酶的序列; 和（d）编码标记的序列，其中所述两个或多个核酸能够彼此进行同源重组，以产生单个核酸，并且其中所述两个或更多个核酸中的至少两个各自包含编码标记的非功能部分的序列; 并且在Rasamsonia细胞中将所述两种或多种核酸彼此重组，并且与靶基因座侧翼的序列重组，使得编码功能性标记的连续核酸序列和编码重组酶的序列插入靶标位点，所述标记编码和/或重组酶编码序列，其侧翼为至少两个位点特异性重组位点，并且所述位点特异性重组位点侧翼为能够与靶基因座侧翼的序列同源重组的序列，从而在 Rasamsonia细胞中的靶标位点。该方法可用于产生无标记的Rasamsonia细胞。

6. 发明授权

EP2825651B1 RECOMBINATION SYSTEM 有权
标题翻译：重组系统
公开(公告)号：EP2825651B1
公开(公告)日：2018-04-25
申请号：EP13708507.2
申请日：2013-03-12
申请人： DSM IP Assets B.V.
发明人： PEIJ, VAN, Noël Nicolaas Maria Elisabeth , BEISHUIZEN, Martina , ARENDSEN, Yvonne Johannes Odilia
IPC分类号： C12N15/63 , C12N15/90 , C12N15/00 , C12N5/00
CPC分类号： C12N15/85 , C12N15/80 , C12N15/81 , C12N15/82 , C12N15/902
摘要： The present invention relates to a method for carrying out recombination at a target locus.

7. 发明公开

EP2825651A1 RECOMBINATION SYSTEM 有权
标题翻译：重组系统
公开(公告)号：EP2825651A1
公开(公告)日：2015-01-21
申请号：EP13708507.2
申请日：2013-03-12
申请人： DSM IP Assets B.V.
发明人： PEIJ, VAN, Noël Nicolaas Maria Elisabeth , BEISHUIZEN, Martina , ARENDSEN, Yvonne Johannes Odilia
IPC分类号： C12N15/63 , C12N15/90 , C12N15/00 , C12N5/00
CPC分类号： C12N15/85 , C12N15/80 , C12N15/81 , C12N15/82 , C12N15/902
摘要： The present invention relates to a method for carrying out recombination at a target locus.
摘要翻译：本发明涉及用于在靶基因座上进行重组的方法，该方法包括：提供两种或更多种核酸，所述两种或更多种核酸一起时包含：（a）能够与靶基因座侧翼序列同源重组的序列; （b）两个或更多个位点特异性重组位点; （c）编码识别位点特异性重组位点的重组酶的序列; 和（d）编码标记的序列，其中所述两种或更多种核酸能够彼此同源重组以产生单个核酸，并且其中所述两种或更多种核酸中的至少两种各自包含编码标记的非功能部分的序列; 并且将所述两种或更多种核酸彼此重组并且与所述靶基因座侧翼的序列重组，从而编码功能性标记的连续核酸序列和编码重组酶的序列被插入靶基因座，所述标记编码和/ 或重组酶编码序列侧翼为至少两个位点特异性重组位点，并且所述位点特异性重组位点侧翼为能够与靶基因座侧翼序列同源重组的序列。

8. 发明公开

EP2825650A1 RECOMBINATION SYSTEM 有权
标题翻译：重组
公开(公告)号：EP2825650A1
公开(公告)日：2015-01-21
申请号：EP13708506.4
申请日：2013-03-12
申请人： DSM IP Assets B.V.
发明人： WIESSENHAAN, Nathalie , KOLEN, Catharina Petronella Antonia Maria , MEIJRINK, Bernard , BOER, Viktor , ROUBOS, Johannes, Andries , ARENDSEN, Yvonne Johannes Odilia
IPC分类号： C12N15/63 , C12N15/90 , C12N15/00 , C12N5/00
CPC分类号： C12N15/102 , C12N15/902 , C12N2800/30
摘要： The present invention relates to a method for carrying out recombination at a target locus.

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