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    • 7. 发明授权
    • Methods for targeted cleavage and recombination of CCR5
    • CCR5靶向切割和重组的方法
    • US08349810B2
    • 2013-01-08
    • US13165866
    • 2011-06-22
    • Jeffrey C. MillerLei Zhang
    • Jeffrey C. MillerLei Zhang
    • A61K48/00C12N5/0783C12N5/074C12N5/0789C12N15/00
    • C12N15/10C07K14/4702C12N9/22
    • Disclosed herein are methods and compositions for targeted cleavage of a genomic sequence, targeted alteration of a genomic sequence, and targeted recombination between a genomic region and an exogenous polynucleotide homologous to the genomic region. The compositions include fusion proteins comprising a cleavage domain (or cleavage half-domain) and an engineered zinc finger domain, as well as polynucleotides encoding same. Fusion proteins comprising cleavage half-domains are used in pairs to reconstitute a functional cleavage domain. In these fusion proteins, the zinc finger domain can be N-terminal to the cleavage half-domain, or the cleavage half-domain can be N-terminal to the zinc finger domain. The availability of fusion endonucleases having these different polarities allows targeting of zinc finger endonucleases either to opposite strands of the DNA target or to the same strand of the DNA target, thereby increasing the number of possible sequences which can be targeted and cleaved by the fusion proteins.
    • 本文公开了用于基因组序列的靶向切割,基因组序列的靶向改变以及基因组区域和与该基因组区域同源的外源多核苷酸之间的靶向重组的方法和组合物。 组合物包括包含切割结构域(或切割半结构域)和工程化锌指结构域的融合蛋白,以及编码它们的多核苷酸。 包含切割半结构域的融合蛋白成对使用以重构功能性切割结构域。 在这些融合蛋白中,锌指结构域可以是切割半结构域的N末端,或者切割半结构域可以是锌指结构域的N末端。 具有这些不同极性的融合核酸内切酶的可获得性允许将锌指内切核酸酶靶向DNA靶的相对链或DNA靶的相同链,从而增加可由融合蛋白靶向和切割的可能序列的数目 。