The cDNA of NCL (GenBank accession no. NM_005381.2) RRM2–4C-terminal domain was amplified and cloned in-frame into the BamHI and EcoRI sites of the eukaryotic expression vector pcDNA3.1(–)myc-his (Invitrogen, Carlsbad, CA, USA) to generate the plasmid pcDNA3.1-NCL (also named pc-NCL). The constructs were confirmed by restriction enzyme digestion along with DNA sequencing analysis.
Short hairpin RNA (shRNA) oligonucleotides against NCL (GenBank accession no: NM_005381.2) were designed by Invitrogen BLOCK-iT™ RNAi Designer and synthesized by TSINGKE, Co. (Wuhan, China). The shRNA oligonucleotides were cloned in-frame into the HindIII and ApaI sites of the pSilencer1.0-U6 (Invitrogen, Carlsbad, CA, USA) to generate the pSilencer1.0-U6-NCL (also named p-sh-NCL). The constructs were confirmed by restriction enzyme digestion along with DNA sequencing analysis. All the DNA preparations were produced using endotoxin-free purification columns (QIAGEN, Valencia, CA, USA).
Bian W.X., Xie Y., Wang X.N., Xu G.H., Fu B.S., Li S., Long G., Zhou X, & Zhang X.L. (2018). Binding of cellular nucleolin with the viral core RNA G-quadruplex structure suppresses HCV replication. Nucleic Acids Research, 47(1), 56-68.
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