title: Efficient and Inducible Use of Artificial MicroRNAs in Marchantia polymorpha.
abstract: We describe the efficient use of artificial microRNAs (amiRs) in Marchantia polymorpha using both endogenous and heterologous primary microRNA (pri-miR) hairpin backbones. Targeting of two transcription factor genes, MpARF1 and MpRR-B, mediating different hormonal responses, demonstrated that amiRs can create specific and reproducible physiological and morphological defects, facilitating interpretation of gene function. A third amiR was designed to target a gene encoding a component of the Polycomb recessive complex 2, MpE(z), and constitutive expression of this amiR results in sporeling lethality. Adaptation of an estrogen-inducible system allowed analysis of the phenotypic effects of induction of this amiR during other stages of the life cycle. We discuss the advantages and challenges of the use of amiRs as a tool for reverse genetic analysis in M. polymorpha.
authors.affiliation: School of Biological Sciences, Monash University, Melbourne, Victoria 3800, Australia.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Flores-Sandoval
authors.affiliation: School of Biological Sciences, Monash University, Melbourne, Victoria 3800, Australia.
authors.forename: Tom
authors.initials: T
authors.lastname: Dierschke
authors.affiliation: School of Biological Sciences, Monash University, Melbourne, Victoria 3800, Australia.
authors.forename: Tom J
authors.initials: TJ
authors.lastname: Fisher
authors.affiliation: School of Biological Sciences, Monash University, Melbourne, Victoria 3800, Australia Department of Plant Biology, UC Davis, Davis, CA 95616, USA john.bowman@monash.edu.
authors.forename: John L
authors.initials: JL
authors.lastname: Bowman
mesh_terms: D001483:Base Sequence; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000378:metabolism; D007210:Indoleacetic Acids / Q000494:pharmacology; D047008:Marchantia / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D010641:Phenotype; D030821:Plants, Genetically Modified; D014170:Transformation, Genetic / Q000187:drug effects
chemical_list: D007210:Indoleacetic Acids; D035683:MicroRNAs
keywords: AUXIN RESPONSE FACTOR; Artificial microRNA; ENHANCER OF ZESTE; Marchantia; MicroRNA; RESPONSE REGULATOR; Reverse genetics