DNA编码动态文库的设计、制备与筛选
Design, preparation, and selection of DNA-encoded dynamic libraries.
作者信息
Li Gang, Zheng Wenlu, Chen Zitian, Zhou Yu, Liu Yu, Yang Junrui, Huang Yanyi, Li Xiaoyu
机构信息
Key Laboratory of Bioorganic Chemistry and Molecular Engineering of the Ministry of Education , Beijing National Laboratory of Molecular Sciences (BNLMS) , College of Chemistry and Molecular Engineering , Peking University , Beijing , China 100871 . Email:
Key Laboratory of Chemical Genomics , School of Chemical Biology and Biotechnology , Peking University Shenzhen Graduate School , Shenzhen , China 518055.
出版信息
Chem Sci. 2015 Dec 1;6(12):7097-7104. doi: 10.1039/c5sc02467f. Epub 2015 Sep 11.
We report a method for the preparation and selection of DNA-encoded dynamic libraries (DEDLs). The library is composed of two sets of DNA-linked small molecules that are under dynamic exchange through DNA hybridization. Addition of the protein target shifted the equilibrium, favouring the assembly of high affinity bivalent binders. Notably, we introduced a novel locking mechanism to stop the dynamic exchange and "freeze" the equilibrium, thereby enabling downstream hit isolation and decoding by PCR amplification and DNA sequencing. Our DEDL approach has circumvented the limitation of library size and realized the analysis and selection of large dynamic libraries. In addition, this method also eliminates the requirement for modified and immobilized target proteins.
我们报道了一种制备和筛选DNA编码动态文库（DEDLs）的方法。该文库由两组通过DNA杂交进行动态交换的DNA连接小分子组成。加入蛋白质靶标会使平衡发生移动，有利于高亲和力二价结合物的组装。值得注意的是，我们引入了一种新型锁定机制来停止动态交换并“冻结”平衡，从而能够通过PCR扩增和DNA测序进行下游命中分离和解码。我们的DEDL方法克服了文库大小的限制，实现了对大型动态文库的分析和筛选。此外，该方法还消除了对修饰和固定化靶蛋白的需求。
Zotero 插件