Cell-internalization SELEX: method for identifying cell-internalizing RNA aptamers for delivering siRNAs to target cells.
Cell-internalization SELEX: method for identifying cell-internalizing RNA aptamers for delivering siRNAs to target cells.
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DOI：
10.1007/978-1-4939-1538-5_11
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发表时间：
2015
期刊：
影响因子：
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通讯作者：
Giangrande, Paloma H
中科院分区：
文献类型：
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作者：
Thiel, William H;Thiel, Kristina W;Flenker, Katie S;Bair, Tom;Dupuy, Adam J;McNamara, James O 2nd;Miller, Francis J;Giangrande, Paloma H
经过十年的努力来解决细胞吸收问题（基于RNA干扰的治疗方法的主要障碍），现在人们对基于RNA干扰的药物重新感到乐观。I期和II期研究已经显示出安全，强大和持久的基因敲除（80- 90%，单次注射后持续一个月）和/或治疗几种肝脏病变的临床益处。尽管前景广阔，但这些研究也强调了对强大的递送技术的需求，以开发用于治疗其他器官系统和疾病的RNAi疗法。siRNA与细胞特异性合成RNA配体（适体）的缀合被提出作为该问题的可行解决方案。虽然令人鼓舞，但RNA适体作为siRNA的递送工具的扩展使用等待能够靶向和进入许多不同细胞类型的细胞质的RNA适体序列的鉴定。我们描述了一种基于细胞的选择过程，用于快速鉴定和表征适合于将siRNA药物递送到靶细胞的细胞质中的RNA适体。这一过程被称为“细胞内化SELEX（指数富集配体系统进化）”，需要多种复杂技术的组合，包括基于细胞培养的SELEX程序、下一代测序（NGS）和新型生物信息学工具。
After a decade of work to address cellular uptake, the principal obstacle to RNAi-based therapeutics, there is now well-deserved, renewed optimism about RNAi-based drugs. Phase I and II studies have shown safe, strong, and durable-gene knockdown (80–90 %, lasting for a month after a single injection) and/or clinical benefit in treating several liver pathologies. Although promising, these studies have also highlighted the need for robust delivery techniques to develop RNAi therapeutics for treating other organ systems and diseases. Conjugation of siRNAs to cell-specific, synthetic RNA ligands (aptamers) is being proposed as a viable solution to this problem. While encouraging, the extended use of RNA aptamers as a delivery tool for siRNAs awaits the identification of RNA aptamer sequences capable of targeting and entering the cytoplasm of many different cell types. We describe a cell-based selection process for the rapid identification and characterization of RNA aptamers suited for delivering siRNA drugs into the cytoplasm of target cells. This process, termed “cell-internalization SELEX (Systematic Evolution of Ligands by Exponential Enrichment),” entails the combination of multiple sophisticated technologies, including cell culture-based SELEX procedures, next-generation sequencing (NGS), and novel bioinformatics tools.