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2018 ; 15
(1
): 35
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CLIP-related methodologies and their application to retrovirology
#MMPMID29716635
Bieniasz PD
; Kutluay SB
Retrovirology
2018[May]; 15
(1
): 35
PMID29716635
show ga
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.
free
free
free
