公开(公告)号：US08883482B2

公开(公告)日：2014-11-11

申请号：US13930663

申请日：2013-06-28

Applicant: Novozymes, Inc.

Inventor： Paul Harris ,  Howard Brody

IPC: C12N15/00 ,  C12N1/15 ,  C12N15/64 ,  C12N15/84

CPC classification number: C12P21/00 ,  C12N15/80 ,  C12N15/902

Abstract: The present invention relates to methods for increasing homologous recombination of a nucleic acid sequence introduced into a host cell, comprising: (a) introducing into a population of filamentous fungal host cells a first nucleic acid sequence encoding a recombination protein and a second nucleic acid sequence comprising one or more regions which are homologous with the genome of the filamentous fungal host cell, wherein (i) the recombination protein promotes the recombination of the one or more regions with the corresponding homologous region in the host's genome to incorporate the second nucleic acid sequence by homologous recombination, and (ii) the number of host cells comprising the incorporated second nucleic acid sequence in the population is increased at least 20% compared to the same population without the first nucleic acid sequence; (b) and isolating from the population a filamentous fungal cell comprising the incorporated second nucleic acid sequence.

Abstract translation: 本发明涉及用于增加引入宿主细胞的核酸序列的同源重组的方法，其包括：（a）向丝状真菌宿主细胞群体中引入编码重组蛋白和第二核酸序列的第一核酸序列 包括与丝状真菌宿主细胞的基因组同源的一个或多个区域，其中（i）重组蛋白促进一个或多个区域与宿主基因组中的相应同源区域的重组，以掺入第二核酸序列 通过同源重组，和（ii）与没有第一核酸序列的相同群体相比，包含所述群体中掺入的第二核酸序列的宿主细胞的数量增加至少20％; （b）并从群体中分离包含掺入的第二核酸序列的丝状真菌细胞。

公开(公告)号：US20140287475A1

公开(公告)日：2014-09-25

申请号：US14255642

申请日：2014-04-17

Applicant: Novozymes A/S ,  Novozymes, Inc.

Inventor： Howard Brody ,  Suchindra Maiyuran ,  Hiroaki Udagawa

IPC: C12P1/02

CPC classification number: C12P1/02 ,  C12N15/113 ,  C12N15/1137 ,  C12N2310/111 ,  C12N2310/14 ,  C12N2310/53

Abstract: The present invention relates to methods for reducing or eliminating the expression of a target gene in a filamentous fungal strain, comprising: (a) inserting into the genome of the filamentous fungal strain a double-stranded transcribable nucleic acid construct comprising a first nucleotide sequence comprising a promoter operably linked to a homologous coding region of the target gene and a second nucleotide sequence comprising the homologous coding region, or a portion thereof, of the target gene, wherein the first and second nucleotide sequences are complementary to each other and the second nucleotide sequence is in reverse orientation relative to the first nucleotide sequence; and (b) inducing production of an interfering RNA encoded by the double-stranded transcribable nucleic acid construct by cultivating the filamentous fungal strain under conditions conducive for production of the interfering RNA; wherein the interfering RNA interacts with RNA transcripts of the target gene to reduce or eliminate expression of the target gene. The present invention also relates to the filamentous fungal strains and to methods of producing a biological substance of interest in such filamentous fungal strains.

Abstract translation: 本发明涉及用于减少或消除丝状真菌菌株中靶基因表达的方法，其包括：（a）将丝状真菌菌株的双链可转录核酸构建体插入基因组中，所述双链可转录核酸构建体包含第一核苷酸序列，其包含 可操作地连接到靶基因的同源编码区的启动子和包含靶基因的同源编码区或其部分的第二核苷酸序列，其中第一和第二核苷酸序列彼此互补，第二核苷酸 序列相对于第一个核苷酸序列处于相反方向; 和（b）通过在有利于产生干扰RNA的条件下培养丝状真菌菌株诱导由双链可转录核酸构建体编码的干扰RNA的产生; 其中所述干扰RNA与所述靶基因的RNA转录物相互作用以减少或消除所述靶基因的表达。 本发明还涉及丝状真菌菌株以及在这种丝状真菌菌株中生产目的生物物质的方法。

公开(公告)号：US20170029821A1

公开(公告)日：2017-02-02

申请号：US15296489

申请日：2016-10-18

Applicant: Novozymes, Inc.

Inventor： Howard Brody ,  Donna Moyer ,  Amanda Fisher

IPC: C12N15/113 ,  C12P1/02

CPC classification number: C12N15/1137 ,  C12N2310/111 ,  C12N2310/14 ,  C12N2310/53 ,  C12P1/02 ,  C12Y302/01037 ,  C12Y604/00

Abstract: The present invention relates to methods of reducing or eliminating expression of a target gene in a filamentous fungal strain by transitive RNA interference.

Abstract translation: 本发明涉及通过传递RNA干扰减少或消除丝状真菌菌株中靶基因表达的方法。

公开(公告)号：US08481320B2

公开(公告)日：2013-07-09

申请号：US13710007

申请日：2012-12-10

Applicant: Novozymes, Inc.

Inventor： Paul Harris ,  Howard Brody

IPC: C12N15/00 ,  C12N1/15 ,  C12N15/64 ,  C12N15/84

CPC classification number: C12P21/00 ,  C12N15/80 ,  C12N15/902

Abstract: The present invention relates to methods for increasing homologous recombination of a nucleic acid sequence introduced into a host cell, comprising: (a) introducing into a population of filamentous fungal host cells a first nucleic acid sequence encoding a recombination protein and a second nucleic acid sequence comprising one or more regions which are homologous with the genome of the filamentous fungal host cell, wherein (i) the recombination protein promotes the recombination of the one or more regions with the corresponding homologous region in the host's genome to incorporate the second nucleic acid sequence by homologous recombination, and (ii) the number of host cells comprising the incorporated second nucleic acid sequence in the population is increased at least 20% compared to the same population without the first nucleic acid sequence; (b) and isolating from the population a filamentous fungal cell comprising the incorporated second nucleic acid sequence.

公开(公告)号：US20150056652A1

公开(公告)日：2015-02-26

申请号：US14536048

申请日：2014-11-07

Applicant: NOVOZYMES, INC.

Inventor： Paul Harris ,  Howard Brody

IPC: C12P21/00

CPC classification number: C12P21/00 ,  C12N15/80 ,  C12N15/902

Abstract: The present invention relates to methods for increasing homologous recombination of a nucleic acid sequence introduced into a host cell, comprising: (a) introducing into a population of filamentous fungal host cells a first nucleic acid sequence encoding a recombination protein and a second nucleic acid sequence comprising one or more regions which are homologous with the genome of the filamentous fungal host cell, wherein (i) the recombination protein promotes the recombination of the one or more regions with the corresponding homologous region in the host's genome to incorporate the second nucleic acid sequence by homologous recombination, and (ii) the number of host cells comprising the incorporated second nucleic acid sequence in the population is increased at least 20% compared to the same population without the first nucleic acid sequence; (b) and isolating from the population a filamentous fungal cell comprising the incorporated second nucleic acid sequence.

Abstract translation: 本发明涉及用于增加引入宿主细胞的核酸序列的同源重组的方法，其包括：（a）向丝状真菌宿主细胞群体中引入编码重组蛋白和第二核酸序列的第一核酸序列 包括与丝状真菌宿主细胞的基因组同源的一个或多个区域，其中（i）重组蛋白促进一个或多个区域与宿主基因组中的相应同源区域的重组，以掺入第二核酸序列 通过同源重组，和（ii）与没有第一核酸序列的相同群体相比，包含所述群体中掺入的第二核酸序列的宿主细胞的数量增加至少20％; （b）并从群体中分离包含掺入的第二核酸序列的丝状真菌细胞。

公开(公告)号：US20130316438A1

公开(公告)日：2013-11-28

申请号：US13930663

申请日：2013-06-28

Applicant: Novozymes, Inc.

Inventor： Paul Harris ,  Howard Brody

IPC: C12N15/90

CPC classification number: C12P21/00 ,  C12N15/80 ,  C12N15/902

Abstract: The present invention relates to methods for increasing homologous recombination of a nucleic acid sequence introduced into a host cell, comprising: (a) introducing into a population of filamentous fungal host cells a first nucleic acid sequence encoding a recombination protein and a second nucleic acid sequence comprising one or more regions which are homologous with the genome of the filamentous fungal host cell, wherein (i) the recombination protein promotes the recombination of the one or more regions with the corresponding homologous region in the host's genome to incorporate the second nucleic acid sequence by homologous recombination, and (ii) the number of host cells comprising the incorporated second nucleic acid sequence in the population is increased at least 20% compared to the same population without the first nucleic acid sequence; (b) and isolating from the population a filamentous fungal cell comprising the incorporated second nucleic acid sequence.