Cell-Internalization SELEX: Method for Identifying Cell-Internalizing RNA Aptamers for Delivering siRNAs to Target Cells

William H Thiel; Kristina W Thiel; Katie S Flenker; Tom Bair; Adam J Dupuy; James O McNamara; Francis J Miller; Paloma H Giangrande

doi:10.1007/978-1-4939-1538-5_11

Book chapter

Cell-Internalization SELEX: Method for Identifying Cell-Internalizing RNA Aptamers for Delivering siRNAs to Target Cells

William H Thiel, Kristina W Thiel, Katie S Flenker, Tom Bair, Adam J Dupuy, James O McNamara, Francis J Miller and Paloma H Giangrande

RNA Interference, pp.187-199

Methods in Molecular Biology, Springer New York

09/23/2014

DOI: 10.1007/978-1-4939-1538-5_11

PMCID: PMC4435695

PMID: 25319652

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Abstract

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.

Cell-internalization SELEX

RNA aptamers

Systematic evolution of ligands by exponential enrichment (SELEX)

Bioinformatics

Next-generation sequencing (NGS)

Quantitative PCR

Cell-targeted aptamers

Details

Title: Subtitle: Cell-Internalization SELEX: Method for Identifying Cell-Internalizing RNA Aptamers for Delivering siRNAs to Target Cells
Creators: William H Thiel - Department of Internal Medicine, University of Iowa, Iowa City, USA
Kristina W Thiel - Department of Internal Medicine, University of Iowa, Iowa City, USA
Katie S Flenker - Department of Internal Medicine, University of Iowa, Iowa City, USA
Tom Bair - Department of Internal Medicine, University of Iowa, Iowa City, USA
Adam J Dupuy - Anatomy & Cell Biology, University of Iowa, Iowa City, USA
James O McNamara - Department of Internal Medicine, University of Iowa, Iowa City, USA
Francis J Miller - Department of Internal Medicine, University of Iowa, Iowa City, USA
Paloma H Giangrande - Department of Radiation Oncology, University of Iowa, Iowa City, USA
Resource Type: Book chapter
Publication Details: RNA Interference, pp.187-199
Series: Methods in Molecular Biology
DOI: 10.1007/978-1-4939-1538-5_11
PMID: 25319652
PMCID: PMC4435695
NLM abbreviation: Methods Mol Biol
eISSN: 1940-6029
ISSN: 1064-3745
Publisher: Springer New York; New York, NY
Language: English
Date published: 09/23/2014
Academic Unit: Anatomy and Cell Biology; Pathology; Cardiovascular Medicine; Radiation Oncology; Obstetrics and Gynecology; Internal Medicine
Record Identifier: 9984025348902771

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