通过基于活细胞的SELEX技术实现细胞类型特异性RNA适体的进化

Evolution of Cell-Type-Specific RNA Aptamers Via Live Cell-Based SELEX.

作者信息

Zhou Jiehua, Rossi John J

机构信息

Division of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, 1500 East Duarte Rd, Duarte, CA, 91010, USA.

Irell and Manella Graduate School of Biological Sciences, Beckman Research Institute of City of Hope, 1500 East Duarte Rd, Duarte, CA, 91010, USA.

出版信息

Methods Mol Biol. 2016;1421:191-214. doi: 10.1007/978-1-4939-3591-8_16.

DOI:10.1007/978-1-4939-3591-8_16

PMID:26965267

Abstract

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 antigen or a particular target cell population. In particular, it offers a facile selection strategy for some special cell-surface proteins that are original glycosylated or heavily post-translationally modified, and are unavailable in their native/active conformation after in vitro expression and purification. In this chapter, we describe evolution of cell-type-specific RNA aptamers targeting the human CCR5 by combining the live cell-based SELEX strategy with high-throughput sequencing (HTS) and bioinformatics analysis.

摘要

基于活细胞的SELEX（指数富集配体系统进化技术）是一种很有前景的方法，用于鉴定能选择性结合细胞表面抗原或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筛选策略，这些蛋白最初是糖基化的或经过大量翻译后修饰，在体外表达和纯化后无法以其天然/活性构象获得。在本章中，我们通过将基于活细胞的SELEX策略与高通量测序（HTS）和生物信息学分析相结合，描述了针对人类CCR5的细胞类型特异性RNA适体的进化。