PAR-CLIP

<p>PAR-CLIP  (photoactivatable ribonucleoside-enhanced crosslinking and immunoprecipitation) is a <a href="/facts/Biochemistry/fa2pWEBp">biochemical</a> method for identifying the binding sites of cellular <a href="/facts/RNA-binding_protein/qTPTczLC">RNA-binding proteins</a> (RBPs) and <a href="/facts/MicroRNA/UjqbYYpe">microRNA</a>-containing ribonucleoprotein complexes (miRNPs). The method relies on the incorporation of <a href="/facts/Ribonucleoside/8lAokLM4">ribonucleoside</a> analogs that are <a href="/facts/Photosensitivity/dohEzKCm">photoreactive</a>, such as 4-thiouridine (4-SU) and 6-thioguanosine (6-SG), into nascent RNA transcripts by living cells. Irradiation of the cells by <a href="/facts/Ultraviolet/kncBUqn5">ultraviolet</a> light of 365 nm <a href="/facts/Wavelength/kcJU0ymz">wavelength</a> induces efficient <a href="/facts/Cross-link/zeQ9hEAA">crosslinking</a> of photoreactive nucleoside–<a href="/facts/Flow_tracer/ssA1CASQ">labeled</a> cellular RNAs to interacting RBPs. <a href="/facts/Immunoprecipitation/PYAYVuz9">Immunoprecipitation</a> of the RBP of interest is followed by isolation of the crosslinked and coimmunoprecipitated RNA. The isolated RNA is converted into a <a href="/facts/Complementary_DNA/P2nWeY4T">cDNA</a> library and is <a href="/facts/Deep_sequencing/NVMzaY0U">deep sequenced</a> using <a href="/facts/Next-generation_sequencing/FjLl8GGa">next-generation sequencing</a> technology.
</p><p>Recently, PAR-CLIP have been applied to determine the transcriptome-wide binding sites of several known RBPs and microRNA-containing ribonucleoprotein complexes at high resolution.
</p>

PAR-CLIP open-in-new

PAR-CLIP