公开(公告)号：US09388417B2

公开(公告)日：2016-07-12

申请号：US14371557

申请日：2013-01-11

Applicant: Korea Advanced Institute of Science and Technology

Inventor： Sang Yup Lee ,  Dokyun Na ,  Seung Min Yoo

IPC: C12N15/63 ,  C12N1/21 ,  C12N15/113 ,  C12P7/22 ,  C12P13/22 ,  C12N15/70 ,  C12P13/00

CPC classification number: C12N15/1138 ,  C12N15/111 ,  C12N15/63 ,  C12N15/67 ,  C12N15/70 ,  C12N2310/11 ,  C12N2310/13 ,  C12N2310/18 ,  C12N2310/3519 ,  C12N2310/531 ,  C12P7/22 ,  C12P13/001 ,  C12P13/225

Abstract: The present invention relates to a novel customized sRNA that reduces gene expression in prokaryotic cells, a preparation method thereof, and the use thereof, and more particularly to a synthetic sRNA comprising an Hfq binding site, derived from the sRNA of any one of MicC, SgrS and MicF, and a region that base-pairs with the target gene mRNA, and to a preparation method thereof and the use thereof. The synthetic sRNA according to the invention has an advantage in that the degree of inhibition of the target gene can be controlled by regulating the ability of the synthetic sRNA to bind to the mRNA of the target gene. The use of the synthetic sRNA that regulates the expression of the target gene makes it possible to effectively construct a recombinant microorganism without using a conventional gene deletion method and to reduce the expression of the target gene, and thus the synthetic sRNA is useful for the production of recombinant microorganisms. Also, the synthetic sRNA can be quickly applied to various strains, and thus is very suitable for the measurement of metabolic capabilities of strains and the selection of the most suitable strain. In addition, recombinant microorganisms, which are obtained by metabolic flux manipulation using the synthetic sRNA and produce tyrosine or cadaverine with high efficiency, are useful in the drug and industrial fields. In other words, the use of the sRNA according to the present invention can make it easy to select target genes whose expression is to be inhibited for the highly efficient production of metabolites. Accordingly, the synthetic sRNA can be used to construct recombinant strains for efficient production of various metabolites and to establish efficient methods for production of various metabolites, and thus is highly useful.

Abstract translation: 本发明涉及减少原核细胞中基因表达的新型定制的sRNA，其制备方法及其用途，更具体地涉及包含衍生自MicC中任一种的sRNA的Hfq结合位点的合成sRNA， SgrS和MicF，以及与靶基因mRNA碱基配对的区域及其制备方法及其用途。 根据本发明的合成sRNA具有以下优点：靶基因的抑制程度可以通过调节合成sRNA结合目标基因的mRNA的能力来控制。 调节靶基因表达的合成sRNA的使用使得可以有效地构建重组微生物而不使用常规的基因缺失方法并降低靶基因的表达，因此合成的sRNA可用于生产 的重组微生物。 此外，合成的sRNA可以快速应用于各种菌株，因此非常适合于测定菌株的代谢能力和选择最合适的菌株。 此外，通过使用合成sRNA的代谢通量操作获得的重组微生物以高效率产生酪氨酸或尸胺，可用于药物和工业领域。 换句话说，根据本发明的sRNA的使用可以使得能够容易地选择其表达被抑制用于代谢物的高效生产的靶基因。 因此，合成的sRNA可用于构建用于有效生产各种代谢物的重组菌株，并建立生产各种代谢物的有效方法，因此是非常有用的。

公开(公告)号：US20170130235A1

公开(公告)日：2017-05-11

申请号：US15317939

申请日：2014-06-11

Applicant: Korea Advanced Institute of Science and Technology

Inventor： Sang Yup Lee ,  Minho Roh ,  Seung Min Yoo

IPC: C12N15/67 ,  C12N15/113

CPC classification number: C12N15/67 ,  C12N15/113 ,  C12N2310/11 ,  C12N2310/111 ,  C12N2310/531

Abstract: The present invention relates to a method for fine-tuning gene expression levels using a synthetic regulatory sRNA in a prokaryotic cell. The present invention can simultaneously, easily, and quickly apply various target gene combinations to various strains without gene deletion through the synthetic regulatory sRNA for regulating gene expression and is therefore very suitable for measuring the metabolizability of each strain and selecting an optimum strain. In addition, the method has the advantages of easily and quickly selecting a target gene for the inhibition of gene expression and expressing the gene thus selected to a desired degree and thus can be used in producing recombinant strains for the efficient production of various metabolites and establishing a method for the efficient production and is therefore very useful.

公开(公告)号：US10196644B2

公开(公告)日：2019-02-05

申请号：US15317939

申请日：2014-06-11

Applicant: Korea Advanced Institute of Science and Technology

Inventor： Sang Yup Lee ,  Minho Roh ,  Seung Min Yoo

IPC: C12N15/67 ,  C12N15/113

Abstract: The present invention relates to a method for fine-tuning gene expression levels using a synthetic regulatory sRNA in a prokaryotic cell. The present invention can simultaneously, easily, and quickly apply various target gene combinations to various strains without gene deletion through the synthetic regulatory sRNA for regulating gene expression and is therefore very suitable for measuring the metabolizability of each strain and selecting an optimum strain. In addition, the method has the advantages of easily and quickly selecting a target gene for the inhibition of gene expression and expressing the gene thus selected to a desired degree and thus can be used in producing recombinant strains for the efficient production of various metabolites and establishing a method for the efficient production and is therefore very useful.

公开(公告)号：US20140377752A1

公开(公告)日：2014-12-25

申请号：US14371557

申请日：2013-01-11

Applicant: Korea Advanced Institute of Science and Technology

Inventor： Sang Yup Lee ,  Dokyun Na ,  Seung Min Yoo

IPC: C12N15/113 ,  C12P13/00 ,  C12N15/70 ,  C12P13/22

CPC classification number: C12N15/1138 ,  C12N15/111 ,  C12N15/63 ,  C12N15/67 ,  C12N15/70 ,  C12N2310/11 ,  C12N2310/13 ,  C12N2310/18 ,  C12N2310/3519 ,  C12N2310/531 ,  C12P7/22 ,  C12P13/001 ,  C12P13/225

Abstract: The present invention relates to a novel customized sRNA that reduces gene expression in prokaryotic cells, a preparation method thereof, and the use thereof, and more particularly to a synthetic sRNA comprising an Hfq binding site, derived from the sRNA of any one of MicC, SgrS and MicF, and a region that base-pairs with the target gene mRNA, and to a preparation method thereof and the use thereof. The synthetic sRNA according to the invention has an advantage in that the degree of inhibition of the target gene can be controlled by regulating the ability of the synthetic sRNA to bind to the mRNA of the target gene. The use of the synthetic sRNA that regulates the expression of the target gene makes it possible to effectively construct a recombinant microorganism without using a conventional gene deletion method and to reduce the expression of the target gene, and thus the synthetic sRNA is useful for the production of recombinant microorganisms. Also, the synthetic sRNA can be quickly applied to various strains, and thus is very suitable for the measurement of metabolic capabilities of strains and the selection of the most suitable strain. In addition, recombinant microorganisms, which are obtained by metabolic flux manipulation using the synthetic sRNA and produce tyrosine or cadaverine with high efficiency, are useful in the drug and industrial fields. In other words, the use of the sRNA according to the present invention can make it easy to select target genes whose expression is to be inhibited for the highly efficient production of metabolites. Accordingly, the synthetic sRNA can be used to construct recombinant strains for efficient production of various metabolites and to establish efficient methods for production of various metabolites, and thus is highly useful.

Abstract translation: 本发明涉及减少原核细胞中基因表达的新型定制的sRNA，其制备方法及其用途，更具体地涉及包含衍生自MicC中任一种的sRNA的Hfq结合位点的合成sRNA， SgrS和MicF，以及与靶基因mRNA碱基配对的区域及其制备方法及其用途。 根据本发明的合成sRNA具有以下优点：靶基因的抑制程度可以通过调节合成sRNA结合目标基因的mRNA的能力来控制。 调节靶基因表达的合成sRNA的使用使得可以有效地构建重组微生物而不使用常规的基因缺失方法并降低靶基因的表达，因此合成的sRNA可用于生产 的重组微生物。 此外，合成的sRNA可以快速应用于各种菌株，因此非常适合于测定菌株的代谢能力和选择最合适的菌株。 此外，通过使用合成sRNA的代谢通量操作获得的重组微生物以高效率产生酪氨酸或尸胺，可用于药物和工业领域。 换句话说，根据本发明的sRNA的使用可以使得能够容易地选择其表达被抑制用于代谢物的高效生产的靶基因。 因此，合成的sRNA可用于构建用于有效生产各种代谢物的重组菌株，并建立生产各种代谢物的有效方法，因此是非常有用的。