期刊
MOLECULAR AND CELLULAR ENDOCRINOLOGY
卷 417, 期 C, 页码 63-72出版社
ELSEVIER IRELAND LTD
DOI: 10.1016/j.mce.2015.09.010
关键词
Luciferase assay; Septo-optic dysplasia; KAL1; Kallmann syndrome; Females
资金
- Birth Defect Foundation-Newlife Project [08-09/29]
- British Society for Paediatric Endocrinology and Diabetes
- Great Ormond Street Hospital Children's Charity
- Wellcome Trust [084361, 086545]
- National Institute for Health Research, Great Ormond Street Hospital for Children/UCL Institute of Child Health Specialist Biomedical Research Centre [11ND01]
- Medical College of Saint Bartholomew's Hospital Trust Charitable Trusts
- National Institute for Health Research, Cambridge Biomedical Research Centre
- Great Ormond Street Hospital Childrens Charity [W1055] Funding Source: researchfish
- Medical Research Council [MR/M000125/1] Funding Source: researchfish
- MRC [MR/M000125/1] Funding Source: UKRI
KAL1 is implicated in 5% of Kallmann syndrome cases, a disorder which genotypically overlaps with septo-optic dysplasia (SOD). To date, a reporter-based assay to assess the functional consequences of KAL1 mutations is lacking. We aimed to develop a luciferase assay for novel application to functional assessment of rare KAL1 mutations detected in a screen of 422 patients with SOD. Quantitative analysis was performed using L6-myoblasts stably expressing FGFR1, transfected with a luciferase-reporter vector containing elements of the FGF-responsive osteocalcin promoter. The two variants assayed [p.K185N, p.P291T], were detected in three females with SOD (presenting with optic nerve hypoplasia, midline and pituitary defects). Our novel assay revealed significant decreases in transcriptional activity [p.K185N: 21% (p < 0.01); p.P291T: 40% (p <0.001)]. Our luciferase-reporter assay, developed for assessment of KAL1 mutations, determined that two variants in females with hypopituitarism/SOD are loss-of-function; demonstrating that this assay is suitable for quantitative assessment of mutations in this gene. (C) 2015 The Authors. Published by Elsevier Ireland Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
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