title: Establishment of HIV-1 model cell line GHOST(3) with stable DRiP78 and NHERF1 knockdown.
abstract: Chemokine receptors CXCR4 and CCR5 are indispensable co-receptors for HIV-1 entry into host cells. In our previous study, we identified that dopamine receptor-interacting protein 78 (DRiP78) and Na(+)-H(+) exchanger regulatory factor 1 (NHERF1) are the CXCR4 and CCR5 homo- or hetero-dimer-interacting proteins. DRiP78 and NHERF1 are able to influence the co-receptor internalization and intracellular trafficking. Over-expression of NHERF1 affects the ligands or HIV-1 gp120-induced CCR5 internalization and HIV-1 production. It is reasonable to speculate that DRiP78 and NHERF1, as well as the signaling pathways involved in viral replication, would probably affect HIV-1 replication through regulating the co-receptors. In this present study, we designed two short hairpin RNAs (shRNAs) targeting the DRiP78 and NHERF1, respectively, and constructed the pLenti6/BLOCK-iT-DEST lentiviral plasmids expressing DRiP78 or NHERF1 shRNA. The packaged lentiviruses were used to transduce the widely-applied HIV-1 model cell line GHOST(3). Then, cells with stable knockdown were established through selecting transduced cells with Blasticidin. This study, for the first time, reported the establishment of the GHOST(3) with DRiP78 and NHERF1 knockdown, which is the first stable cell line with HIV-1 co-receptor-interacting molecular defects.
authors.affiliation: Center for Translational Medicine, Huaihe Clinical Institute, Henan University, Kaifeng 475000, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Guangdong Institute of Public Health and Institute of AIDS Control and Prevention, Guangdong Center for Disease Control and Prevention, Guangzhou 510433, China.
authors.forename: Xu-He
authors.initials: XH
authors.lastname: Huang
authors.affiliation: Guangdong Institute of Public Health and Institute of AIDS Control and Prevention, Guangdong Center for Disease Control and Prevention, Guangzhou 510433, China.
authors.forename: Ping-Ping
authors.initials: PP
authors.lastname: Zhou
authors.affiliation: Guangdong Institute of Public Health and Institute of AIDS Control and Prevention, Guangdong Center for Disease Control and Prevention, Guangzhou 510433, China.
authors.forename: Guo-Long
authors.initials: GL
authors.lastname: Yu
authors.affiliation: Guangdong Institute of Public Health and Institute of AIDS Control and Prevention, Guangdong Center for Disease Control and Prevention, Guangzhou 510433, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Yan
authors.affiliation: Guangdong Institute of Public Health and Institute of AIDS Control and Prevention, Guangdong Center for Disease Control and Prevention, Guangzhou 510433, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Qin
authors.affiliation: Guangdong Institute of Public Health and Institute of AIDS Control and Prevention, Guangdong Center for Disease Control and Prevention, Guangzhou 510433, China.
authors.forename: Xin-Ge
authors.initials: XG
authors.lastname: Yan
authors.affiliation: Guangdong Institute of Public Health and Institute of AIDS Control and Prevention, Guangdong Center for Disease Control and Prevention, Guangzhou 510433, China.
authors.forename: Li-Mei
authors.initials: LM
authors.lastname: Diao
authors.affiliation: Guangdong Institute of Public Health and Institute of AIDS Control and Prevention, Guangdong Center for Disease Control and Prevention, Guangzhou 510433, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Lin
authors.affiliation: Center for Translational Medicine, Huaihe Clinical Institute, Henan University, Kaifeng 475000, China;Guangdong Institute of Public Health and Institute of AIDS Control and Prevention, Guangdong Center for Disease Control and Prevention, Guangzhou 510433, China. yqkuang@henu.edu.cn.
authors.forename: Yi-Qun
authors.initials: YQ
authors.lastname: Kuang
mesh_terms: D002460:Cell Line / Q000378:metabolism / Q000821:virology; D005326:Fetal Proteins / Q000235:genetics* / Q000378:metabolism; D055785:Gene Knockdown Techniques; D015658:HIV Infections / Q000235:genetics* / Q000378:metabolism / Q000821:virology*; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D018832:Molecular Chaperones / Q000235:genetics* / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D017923:Sodium-Hydrogen Exchangers / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication
chemical_list: C476535:DNAJC14 protein, human; D005326:Fetal Proteins; D018832:Molecular Chaperones; D010750:Phosphoproteins; D034741:RNA, Small Interfering; D017923:Sodium-Hydrogen Exchangers; C093376:sodium-hydrogen exchanger regulatory factor
keywords: DRiP78; GHOST(3) cells; HIV-1; NHERF1; shRNA