Efficient and inducible use of artificial microRNAs in Marchantia polymorpha

Research output: Contribution to journalArticleResearchpeer-review

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.
Original languageEnglish
Pages (from-to)281-290
Number of pages10
JournalPlant and Cell Physiology
Volume57
Issue number2
DOIs
Publication statusPublished - 2016

Keywords

  • Artificial microRNA
  • AUXIN RESPONSE FACTOR
  • ENHANCER OF ZESTE
  • Marchantia
  • MicroRNA
  • RESPONSE REGULATOR
  • Reverse genetics

Cite this

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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.",
keywords = "Artificial microRNA, AUXIN RESPONSE FACTOR, ENHANCER OF ZESTE, Marchantia, MicroRNA, RESPONSE REGULATOR, Reverse genetics",
author = "Eduardo Flores-Sandoval and Tom Dierschke and Fisher, {Tom J} and Bowman, {John L}",
year = "2016",
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language = "English",
volume = "57",
pages = "281--290",
journal = "Plant and Cell Physiology",
issn = "0032-0781",
publisher = "Oxford University Press",
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}

Efficient and inducible use of artificial microRNAs in Marchantia polymorpha. / Flores-Sandoval, Eduardo; Dierschke, Tom; Fisher, Tom J; Bowman, John L.

In: Plant and Cell Physiology, Vol. 57, No. 2, 2016, p. 281-290.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Efficient and inducible use of artificial microRNAs in Marchantia polymorpha

AU - Flores-Sandoval, Eduardo

AU - Dierschke, Tom

AU - Fisher, Tom J

AU - Bowman, John L

PY - 2016

Y1 - 2016

N2 - 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.

AB - 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.

KW - Artificial microRNA

KW - AUXIN RESPONSE FACTOR

KW - ENHANCER OF ZESTE

KW - Marchantia

KW - MicroRNA

KW - RESPONSE REGULATOR

KW - Reverse genetics

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U2 - 10.1093/pcp/pcv068

DO - 10.1093/pcp/pcv068

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SP - 281

EP - 290

JO - Plant and Cell Physiology

JF - Plant and Cell Physiology

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