会员体验
专利管家(专利管理)
工作空间(专利管理)
风险监控(情报监控)
数据分析(专利分析)
侵权分析(诉讼无效)
联系我们
交流群
官方交流:
QQ群: 891211   
微信请扫码    >>>
现在联系顾问~
热词
    • 1. 发明申请
    • METHOD FOR HOMOLOGOUS RECOMBINATION
    • 同种异体重建方法
    • US20140154807A1
    • 2014-06-05
    • US14146858
    • 2014-01-03
    • Hesselien TOUW-RIELMarco Alexander BERG VAN DEN
    • Hesselien TOUW-RIELMarco Alexander BERG VAN DEN
    • C12N15/80
    • C12N15/80C07K14/385C12N15/1058C12N15/902C12N2800/30
    • The present invention discloses a method to construct eukaryotic cells having a target sequence in a chromosomal DNA sequence replaced by a replacement sequence of interest comprising: modifying a parent eukaryotic cell with a preference for NHR to provide a eukaryotic cell having an increased HR/NHR ratio as compared to the parent cell, providing two sets of DNA molecules of which the first set comprises DNA molecules each comprising a first non-functional fragment of the replacement sequence of interest flanked at its 5′-side by a DNA sequence substantially homologous to a sequence of the chromosomal DNA flanking the target sequence and the second set comprises DNA molecules each comprising a second non-functional fragment of the DNA replacement sequence of interest overlapping with the first non-functional fragment and flanked at its 3′-side by a DNA sequence substantially homologous to a sequence of the chromosomal DNA flanking the target sequence, wherein the first and second non-functional fragments become functional upon recombination; transforming the modified eukaryotic cells with both sets of DNA molecules; and growing the cells to obtain transformed progeny cells having a DNA molecule from the first set recombined with a DNA molecule from the second set into a DNA molecule comprising a functional replacement sequence and having the target sequence replaced by the functional replacement sequence.
    • 本发明公开了一种构建真核细胞的方法,所述真核细胞在染色体DNA序列中被替换的目标序列替代,所述染色体DNA序列被替换的目标序列所替代,包括:以优选NHR修饰亲本真核细胞以提供具有增加的HR / NHR比的真核细胞 与亲本细胞相比,提供两组DNA分子,其中第一组包含DNA分子,每个DNA分子包含在其5'侧两侧的替换序列的第一个非功能性片段,其基本上与 在靶序列侧翼的染色体DNA的序列和第二组包含DNA分子,每个DNA分子包含与第一非功能片段重叠的DNA置换序列的第二个非功能性片段,并在其3'侧通过DNA侧接 基本上与靶序列侧翼的染色体DNA的序列同源的序列,其中第一和第二 非功能性片段在重组时变得有功能; 用两组DNA分子转化修饰的真核细胞; 并生长细胞以获得具有来自第一组的DNA分子的转化后代细胞与来自第二组的DNA分子重组为包含功能性替换序列的DNA分子并且将靶序列替换为功能性替换序列。