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Plants: miRNA and RNA silencing

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Plants: miRNA and RNA silencing. pre-miRNA structure. miRNA. biogenesis. small RNA biogenesis. miRNA in plants: developmental role. miRNA siRNA in action ... – PowerPoint PPT presentation

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Title: Plants: miRNA and RNA silencing


1
Plants miRNA and RNA silencing
2
pre-miRNA structure
3
miRNAbiogenesis
4
small RNA biogenesis
5
miRNA in plantsdevelopmental role
6
miRNA siRNA in action
7
RdDM RNA-directed DNA Methylation
8
miRNA research tools
  • biochemical approach
  • isolation, RT, PCR, concatamerization, sequencing
  • microarrays (tissue specificity)
  • bioinformatic alignment studies
  • grafting experiments
  • A. tumefaciens infiltration assay
  • wheat germ in vitro assay
  • in vivo cleavage product detection

9
miRNA research tools
  • biochemical approach
  • bioinformatic alignment studies
  • high complementarity to target mRNAs in plants
  • needs experimental confirmation
  • grafting experiments
  • A. tumefaciens infiltration assay
  • wheat germ in vitro assay
  • in vivo cleavage product detection

10
miRNA research tools
  • biochemical approach
  • bioinformatic alignment studies
  • grafting experiments
  • intercellular RNA signaling through PD phloem
  • A. tumefaciens infiltration assay
  • wheat germ in vitro assay
  • in vivo cleavage product detection

11
miRNA research tools
  • biochemical approach
  • bioinformatic alignment studies
  • grafting experiments
  • A. tumefaciens infiltration assay
  • binary shRNA vectors available
  • constitutive knockdown of target genes
  • wheat germ in vitro assay
  • in vivo cleavage product detection

12
miRNA research tools
  • biochemical approach
  • bioinformatic alignment studies
  • grafting experiments
  • A. tumefaciens infiltration assay
  • wheat germ in vitro assay
  • contains DCL, RdRP
  • in vivo cleavage product detection

13
miRNA research tools
  • biochemical approach
  • bioinformatic alignment studies
  • grafting experiments
  • A. tumefaciens infiltration assay
  • wheat germ in vitro assay
  • in vivo cleavage product detection
  • sensor technology

14
Role and conservation of Arabidopsis miRNA
15
miRNAs important for plant development
16
effects of miRNA156 overexpression
17
miRNAs important for plant development
18
effects of miRNA164b overexpression
19
miRNA siRNA application
Genscript RNAi vector
20
miRNA siRNA application
  • research
  • gene knockdown
  • transient by siRNA infiltration
  • constitutive by shRNA binary vectors
  • tissue-specific by adequate promoters
  • chromatin-based transcriptional silencing
  • promoter silencing
  • transposon silencing
  • agriculture
  • adapt plant mechanism to combat viral infections
  • silencing in related species (eg. male sterility
    in crops)
  • BASF plant nonhost disease resistance screen
  • medicine
  • gene therapy siRNA drugs
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