title: In-vivo quantification of primary microRNA processing by Drosha with a luciferase based system.
abstract: The RNAse III Drosha is responsible for the first step of microRNA maturation, the cleavage of primary miRNA to produce the precursor miRNA. Processing by Drosha is finely regulated and influences the amount of mature microRNA in a cell. We describe in the present work a method to quantify Drosha processing activity in-vivo, which is applicable to any microRNA. With respect to other methods for measuring Drosha activity, our system is faster and scalable, can be used with any cellular system and does not require cell sorting or use of radioactive isotopes. This system is useful to study regulation of Drosha activity in physiological and pathological conditions.
authors.affiliation: Department of Internal Medicine III, University Hospital Ulm, Albert-Einstein-Allee 23, 89081 Ulm, Germany.
authors.forename: Danilo
authors.initials: D
authors.lastname: Allegra
authors.forename: Daniel
authors.initials: D
authors.lastname: Mertens
mesh_terms: D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D008722:Methods; D035683:MicroRNAs / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D043244:Ribonuclease III / Q000235:genetics / Q000378:metabolism*
chemical_list: D035683:MicroRNAs; D008156:Luciferases; D043244:Ribonuclease III