Evolution of Cell-Type-Specific RNA Aptamers via Live Cell-Based SELEX.
Evolution of Cell-Type-Specific RNA Aptamers via Live Cell-Based SELEX.
复制标题
通过基于活细胞的 SELEX 进化细胞类型特异性 RNA 适体。
DOI：
10.1007/978-1-0716-3191-1_22
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Rossi,JohnJ
中科院分区：
文献类型：
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作者：
Herrera,Alberto;Zhou,Jiehua;Song,Min-Sun;Rossi,JohnJ
基于活细胞的SELEX(系统进化配体指数富集法)是一种很有前途的方法，可以识别能够选择性地与细胞表面受体结合或识别特定靶细胞群体的适体。特别是，它为一些特殊的细胞表面蛋白提供了一种简便的选择策略，这些蛋白最初是糖基化的或严重的翻译后修饰，但在体外表达和纯化后无法获得其天然/活性构象。在本章中，我们描述了一种通用的程序，通过结合基于活细胞的SELEX策略与高通量测序(HTS)和生物信息学分析，针对细胞膜结合靶标的细胞类型特异性RNA适体的进化。
Live cell-based SELEX (Systematic Evolution of Ligand EXponential enrichment) is a promising approach for identifying aptamers that can selectively bind to a cell-surface receptor or recognize a particular target cell population. In particular, it offers a facile selection strategy for some special cell-surface proteins that are originally glycosylated or heavily posttranslationally modified and are unavailable in their native/active conformation after in vitro expression and purification. In this chapter, we describe a generalized procedure for evolution of cell type-specific RNA aptamers targeting a cell membrane bound target by combining the live cell-based SELEX strategy with high-throughput sequencing (HTS) and bioinformatics analysis.