Volume 36 Issue 3
May  2015
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Lin ZHANG, Xu-He HUANG, Ping-Ping ZHOU, Guo-Long YU, Jin YAN, Bing QIN, Xin-Ge YAN, Li-Mei DIAO, Peng LIN, Yi-Qun KUANG. Establishment of HIV-1 model cell line GHOST(3) with stable DRiP78 and NHERF1 knockdown. Zoological Research, 2015, 36(3): 161-166.
Citation: Lin ZHANG, Xu-He HUANG, Ping-Ping ZHOU, Guo-Long YU, Jin YAN, Bing QIN, Xin-Ge YAN, Li-Mei DIAO, Peng LIN, Yi-Qun KUANG. Establishment of HIV-1 model cell line GHOST(3) with stable DRiP78 and NHERF1 knockdown. Zoological Research, 2015, 36(3): 161-166.

Establishment of HIV-1 model cell line GHOST(3) with stable DRiP78 and NHERF1 knockdown

Funds:  This study was supported by the Science and Technology Planning Project of Guangdong Province of China (2012B031800267); the Natural Science Foundation of Guangdong Province of China (S2013010011860) and the National Natural Science Foundation of China (31200130 and 81371812)
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  • Corresponding author: Yi-Qun KUANG
  • Received Date: 2015-01-21
  • Rev Recd Date: 2015-03-20
  • Publish Date: 2015-05-08
  • 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.
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