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- W3028668028 abstract "ObjectiveTo investigate the effect of cone-rod homeobox containing gene (CRX) mutation on the transcriptional expression of the rhodopsin gene and the stability of the neural retina leucine zipper (NRL) protein, both in the presence or absence of the NRL gene.MethodsIn this experimental study, different expression vectors carrying the wild-type (WT) or the c. C766T (p.Q256X) nonsense mutant CRX were constructed with the NRL and the bovine rhodopsin gene promoter sub-luciferase (pBR130-luc). The vectors were then transfected into 293T and ARPE-19 cell lines, respectively. Then, dual-luciferase assays and Western blotting analyses were performed. The housekeeping protein glyceraldehyde-3-phosphate dehydrogenase was used as an internal reference marker.ResultsThe expression vector of CRX/WT caused a 5-fold increased activation of bovine rhodopsin promoter compared to a 25% decrease by CRX/c.C766T. Co-expression of CRX/WT and NRL increased activation by 30-fold compared to a 7.7% decrease by the co-expression of CRX/c.C766T and NRL. Also, expression of NRL protein increased with the co-expression of CRX/c.C766T and NRL.ConclusionThe in vitro experiments suggest that the c.C766T (p. Q256X) mutant of the CRX gene down-regulated the transcriptional expression of rhodopsin and interfered with the homeostasis of CRX and NRL proteins.Key words: Cone-rod homeobox containing gene; Neural retina leucine zipper; Cone-rod dystrophy 2; Hereditary retinal degeneration; Dual-luciferase assay; Western blotting" @default.
- W3028668028 created "2020-06-05" @default.
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- W3028668028 date "2017-01-25" @default.
- W3028668028 modified "2023-09-23" @default.
- W3028668028 title "Hereditary retinal degeneration associated with CRX gene mutation interference with the transcriptional expression of rhodopsin and NRL stability" @default.
- W3028668028 doi "https://doi.org/10.3760/cma.j.issn.1674-845x.2017.01.009" @default.
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