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Lariat aptamer: aptamer candidate exclusion by nuclease digestion

机译:套索适体:通过核酸酶消化排除适体候选者

摘要

A method for identifying aptamers that bind to target molecules may include contacting an oligonucleotide library with target molecule and digesting unbound oligonucleotides with one or more endonucleases, one or more exonucleases, or one or more endonucleases in combination with one or more exonucleases. The unbound oligonucletides lend themselves to nuclease digestion because their flanking sequences are complementary and form predictable, terminal, secondary structure (a Lariat Aptamer). Bound molecules (aptamers) are protected from nuclease digestion and become enriched. A method for identifying aptamers may further include optionally subjecting selected aptamers to one or more rounds of selection under conditions of increased stringency. A method for identifying aptamers may include yet further amplifying selected aptamers. The described methods may be performed in a screen for identifying aptamers either alone or in combination with other methods typically employed in the art for selecting aptamers (such as, e.g., SELEX). Also contemplated herein are systems and kits for accomplishing the above.
机译:鉴定与靶分子结合的适体的方法可包括使寡核苷酸文库与靶分子接触,并用一种​​或多种核酸内切酶,一种或多种核酸外切酶或与一种或多种核酸外切酶组合的一种或多种核酸内切酶消化未结合的寡核苷酸。未结合的寡核苷酸使自己适合核酸酶消化,因为它们的侧翼序列是互补的,并形成可预测的末端二级结构(Lariat Aptamer)。结合的分子(适体)受到保护,不被核酸酶消化并富集。鉴定适体的方法可以进一步包括在严格性提高的条件下任选地使选择的适体经历一轮或多轮选择。鉴定适体的方法可以包括进一步扩增选择的适体。可以在用于识别适体的屏幕中单独或与本领域中通常用于选择适体的其他方法(例如,SELEX)结合使用所述方法。本文还考虑了用于实现上述目的的系统和套件。

著录项

  • 公开/公告号US8680017B2

    专利类型

  • 公开/公告日2014-03-25

    原文格式PDF

  • 申请/专利权人 GREGORY FRANCIS LOPREATO;

    申请/专利号US201113026244

  • 发明设计人 GREGORY FRANCIS LOPREATO;

    申请日2011-02-12

  • 分类号C40B30/04;

  • 国家 US

  • 入库时间 2022-08-21 16:01:25

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