title: Efficient use of artificial micro-RNA to downregulate the expression of genes at the post-transcriptional level in Arabidopsis thaliana.
abstract: Micro-RNAs are cellular components regulating gene expression at the post-transcription level. In the present study, artificial micro-RNAs were used to decrease the transcript level of two genes, AtExpA8 (encoding an expansin) and AHL25 (encoding an AT-hook motif nuclear localized protein) in Arabidopsis thaliana. The backbone of the Arabidopsis endogenous MIR319a micro-RNA was used in a site-directed mutagenesis approach for the generation of artificial micro-RNAs targeting two genes. The recombinant cassettes were expressed under the control of the CaMV 35S promoter in individual A. thaliana plants. Transgenic lines of the third generation were tested by isolating total RNA and by subsequent cDNA synthesis using oligo-dT18 primers and mRNAs as templates. The expression of the two target genes was checked through quantitative real-time polymerase chain reaction to confirm reduced transcript levels for AtExpA8 and AHL25. Downregulation of AtExpA8 resulted in the formation of short hypocotyls compared with those of the wild-type control in response to low pH and high salt concentration. This technology could be used to prevent the expression of exogenous and invading genes posing a threat to the normal cellular physiology of the host plant.
authors.affiliation: Department of Genetics, Garden Campus, Hazara University, Mansehra, Pakistan.|Institute of Biochemistry and Biology, University of Potsdam, Potsdam, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Ud-Din
authors.affiliation: Institute of Biochemistry and Biology, University of Potsdam, Potsdam, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Rauf
authors.affiliation: Department of Genetics, Garden Campus, Hazara University, Mansehra, Pakistan.
authors.forename: S
authors.initials: S
authors.lastname: Ghafoor
authors.affiliation: Department of Zoology, Garden Campus, Hazara University, Mansehra, Pakistan.
authors.forename: M N K
authors.initials: MN
authors.lastname: Khattak
authors.affiliation: Dr. Panjwani Center for Molecular Medicine and Drug Research, University of Karachi, Karachi, Pakistan.
authors.forename: M W
authors.initials: MW
authors.lastname: Hameed
authors.affiliation: National Agricultural Research Centre, Islamabad, Pakistan.
authors.forename: H
authors.initials: H
authors.lastname: Shah
authors.affiliation: Department of Genetics, Garden Campus, Hazara University, Mansehra, Pakistan.
authors.forename: S
authors.initials: S
authors.lastname: Jan
authors.affiliation: Department of Genetics, Garden Campus, Hazara University, Mansehra, Pakistan.
authors.forename: K
authors.initials: K
authors.lastname: Muhammad
authors.affiliation: Department of Zoology, Garden Campus, Hazara University, Mansehra, Pakistan.
authors.forename: A
authors.initials: A
authors.lastname: Rehman
authors.affiliation: Department of Genetics, Garden Campus, Hazara University, Mansehra, Pakistan.
authors.lastname: Inamullah
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D015536:Down-Regulation; D018506:Gene Expression Regulation, Plant; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: C000595631:MIRN319 microRNA, Arabidopsis; D035683:MicroRNAs; D010940:Plant Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; C095825:expansin protein, plant