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P4‐144: A case of familial CJD with a PRNP E200K mutation
Author(s) -
Kwon Yooseok,
Yang Youngsoon,
Kim SangYun
Publication year - 2012
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2012.05.1847
Subject(s) - dysarthria , mutation , medicine , prnp , prion protein , pediatrics , audiology , pathology , disease , genetics , biology , gene
of MAPT sequence including 5 kb of downstream regulatory sequence. In this study, we cloned the 4.1 kb MAPT 3’ UTR from the H1c and H2 PACs into vectors (pmirGLO, Promega) in which the luciferase gene is fused the MAPT 3’ UTR (luc-H1-UTR and luc-H2-UTR). In parallel, we performed an in silico analysis of the MAPT 3’ UTR to identify potential miRNA regulators of MAPT expression. Candidate miRNAs were then cloned into pCMVMIR (Origene) and co-transfected with luciferase reporter constructs into human neuroblastoma cell culture models. Results: Eleven candidate miRNAs predicted to bind the MAPT 3’ UTR were identified by selecting candidates predicted by both TargetScan and PicTa and were then cloned into pCMV-mir. Subsequent co-transfection of miRNA vectors with the luciferase reporter vectors containing the MAPT 3’ UTRs identified 3 miRNAs that down regulated luciferase expression. Luc-H1-UTR and Luc-H2-UTR displayed a > 50% reduction in luciferase activity with one candidate (P< 0.001). Moreover, Luc-H2-UTR demonstrated w35% less luciferase activity with two additional candidates (P< 0.001) which had no significant effect on the Luc-H1-UTR vectors. Conclusions: These findings suggest a role for miRNA-mediated regulation of expression at the MAPT locus. Furthermore, this regulation may be subject to the presence of specific MAPT haplotype polymorphisms.