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Characterisation of RPGRIP1 and its interacting partners in Zebrafish
Author(s) -
KOTAPATI RAGHUPATHY R,
MCCULLOCH D,
CRAFT J,
SHU X
Publication year - 2014
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2014.f016.x
Subject(s) - zebrafish , biology , pax6 , luciferase , gene , transcription (linguistics) , transcription factor , computational biology , reporter gene , gene expression , microbiology and biotechnology , genetics , transfection , linguistics , philosophy
Purpose Mutations in the gene RPGRIP1 causes LCA, most severe form of retinal dystrophy. Several studies using animal models demonstrate RPGRIP1 show species specific expression and different isoforms of the gene have subcellular specific localisation. RPGRIP1 is mainly localised in the connecting cilium and interacts with RPGR, RPGRIP1L, but the exact function of the RPGRIP1 is still unclear. Methods We have carried out gene expression pattern analysis at various stages of development and different adult tissues of zebrafish using PCR and SYBR green based QPCR. We have also carried promoter analysis of human and zebrafish RPGRIP1 to understand mechanism regulating the transcription of these genes in retina using dual luciferase reporter assay. Results The expression pattern analysis of ZRPGR demonstrates ZRPGR2ORF15 and ZRPGR2Ex1‐17 have different expression pattern in the tissues and the developmental stages. ZRPGR2ORF15 is highly expressed in the eye and brain compared to the expression of ZRPGR2Ex1‐17. Both ZRPGRIP1 and ZRPGRIP1L show higher expression in the eye and ZRPGRIP1Like is highly expressed during the early stages of the development. The binding sites of key transcription binding factors such as CRX, NRL, PAX6 were identified in the 3Kb region upstream of the human RPGRIP1start‐coding site, and a 288bp fragment upstream of transcription start site directed higher luciferase activity in the RPE‐1 cells. Conclusion In this study we found that ZRPGRIP1 is highly expressed in the eye and in the early stages of development than that of its interacting partners ZRPGR and ZRPGRIP1L. 288bp promoter region upstream of human RPGRIP1 transcription start site showed high luciferase activity and supposed to be the core promoter of RPGRIP1.