title: Silencing of Parkinson's disease-associated genes with artificial mirtron mimics of miR-1224.
abstract: Mirtrons are a recently described category of microRNA (miRNA) relying on splicing rather than processing by the microprocessor complex to generate pre-miRNA precursors of the RNA interference (RNAi) pathway. Their discovery and subsequent verification provides important information about a distinct class of miRNA and inherent advantages that could be exploited to silence genes of interest. These include micro-processor-independent biogenesis, pol-II-dependent transcription, accurate species generation and the delivery of multiple artificial mirtrons as introns within a single host transcript. Here we determined the sequence motifs required for correct processing of the mmu-miR-1224 mirtron and incorporated these into artificial mirtrons targeting Parkinson's disease-associated LRRK2 and α-synuclein genes. By incorporating these rules associated with processing and splicing, artificial mirtrons could be designed and made to silence complementary targets either at the mRNA or protein level. We further demonstrate with a LRRK2 targeting artificial mirtron that neuronal-specific silencing can be directed under the control of the human synapsin promoter. Finally, multiple mirtrons were co-delivered within a single host transcript, an eGFP reporter, to allow simultaneous targeting of two or more targets in a combinatorial approach. Thus, the unique characteristics of artificial mirtrons make this an attractive approach for future RNAi applications.
authors: Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
authors: Christopher R
authors: CR
authors: Sibley
authors: Yiqi
authors: Y
authors: Seow
authors: Helen
authors: H
authors: Curtis
authors: Marc S
authors: MS
authors: Weinberg
authors: Matthew J A
authors: MJ
authors: Wood
mesh_terms: D002460:Cell Line; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D035683:MicroRNAs / Q000737:chemistry*; D018716:Molecular Mimicry; D009474:Neurons / Q000378:metabolism; D010300:Parkinson Disease / Q000235:genetics; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D034622:RNA Interference; D012326:RNA Splicing; D012333:RNA, Messenger / Q000737:chemistry; D051844:alpha-Synuclein / Q000235:genetics*
chemical_list: C558413:MIRN1224 microRNA, mouse; D035683:MicroRNAs; D012333:RNA, Messenger; D051844:alpha-Synuclein; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases