CLIP-related methodologies and their application to retrovirology
Open Access
- 2 May 2018
- journal article
- review article
- Published by Springer Science and Business Media LLC in Retrovirology
- Vol. 15 (1), 35
- https://doi.org/10.1186/s12977-018-0417-2
Abstract
Virtually every step of HIV-1 replication and numerous cellular antiviral defense mechanisms are regulated by the binding of a viral or cellular RNA-binding protein (RBP) to distinct sequence or structural elements on HIV-1 RNAs. Until recently, these protein–RNA interactions were studied largely by in vitro binding assays complemented with genetics approaches. However, these methods are highly limited in the identification of the relevant targets of RBPs in physiologically relevant settings. Development of crosslinking-immunoprecipitation sequencing (CLIP) methodology has revolutionized the analysis of protein–nucleic acid complexes. CLIP combines immunoprecipitation of covalently crosslinked protein–RNA complexes with high-throughput sequencing, providing a global account of RNA sequences bound by a RBP of interest in cells (or virions) at near-nucleotide resolution. Numerous variants of the CLIP protocol have recently been developed, some with major improvements over the original. Herein, we briefly review these methodologies and give examples of how CLIP has been successfully applied to retrovirology research.Keywords
Funding Information
- National Institute of General Medical Sciences (GM122458, U54 GM103297)
- National Institute of Allergy and Infectious Diseases (AI50111)
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