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Use of transposable elements for altering gene expression

机译:转座因子用于改变基因表达的用途

摘要

Methods of repressing expression of a recombinant gene in a cell are provided. The methods include the steps of introducing a transposase DNA binding motif into or adjacent to the gene, and introducing into the cell a transposase that is capable of binding to the transposase DNA binding motif. Methods of producing a population of cells of an organism that vary in their expression of a gene are also provided. The methods involve transfecting the cells with a first polynucleotide sequence encoding the target gene operably linked to a promoter such that the target gene is expressed in the cells, wherein the vector has at least one transposase DNA binding motif within or adjacent to the target gene; and transfecting some of the cells with a second polynucleotide encoding the transposable element operably linked to a second promoter such that the transposable element is expressed in the cells. In these methods, the target gene in the cells transfected with the second polynucleotide exhibits reduced expression when compared to the target gene in the cells that are not transfected with the second polynucleotide. Kits having a first polynucleotide with a gene and a DNA transposase binding motif in or adjacent to the gene, and a second polynucleotide comprising a gene encoding a transposase that is capable of binding to the transposase DNA binding motif are also provided.
机译:提供了抑制重组基因在细胞中表达的方法。该方法包括以下步骤:将转座酶DNA结合基序引入基因中或与基因相邻,以及将能够结合转座酶DNA结合基序的转座酶引入细胞。还提供了产生其基因表达变化的生物体细胞群体的方法。该方法包括用编码与启动子可操作地连接的靶基因的第一多核苷酸序列转染细胞,以使靶基因在细胞中表达,其中载体在靶基因内或附近具有至少一个转座酶DNA结合基序。然后用编码与第二启动子可操作连接的转座因子的第二多核苷酸转染一些细胞,从而使转座因子在细胞中表达。在这些方法中,与未用第二多核苷酸转染的细胞中的靶基因相比,用第二多核苷酸转染的细胞中的靶基因表现出降低的表达。还提供了具有第一多核苷酸的试剂盒,该第一多核苷酸具有一个基因以及在该基因中或与该基因相邻的DNA转座酶结合基序,以及第二多核苷酸,该第二多核苷酸包含编码能够结合该转座酶DNA结合基序的转座酶的基因。

著录项

  • 公开/公告号US7064246B2

    专利类型

  • 公开/公告日2006-06-20

    原文格式PDF

  • 申请/专利权人 AMY F. MACRAE;

    申请/专利号US20020138221

  • 发明设计人 AMY F. MACRAE;

    申请日2002-05-01

  • 分类号C12N15/82;

  • 国家 US

  • 入库时间 2022-08-21 21:43:16

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