A modular steroid-inducible gene expression system for use in rice.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
Read Online Read More Add to Saved list
  • Additional Information
    • Source:
      Publisher: BioMed Central Country of Publication: England NLM ID: 100967807 Publication Model: Electronic Cited Medium: Internet ISSN: 1471-2229 (Electronic) Linking ISSN: 14712229 NLM ISO Abbreviation: BMC Plant Biol Subsets: MEDLINE
    • Publication Information:
      Original Publication: London : BioMed Central, [2001-
    • Subject Terms:
      Gene Expression*; Dexamethasone/*administration & dosage ; Gene Expression Profiling/*methods ; Glucuronidase/*genetics ; Oryza/*genetics ; Plant Proteins/*genetics; Gene Regulatory Networks/drug effects ; Genes, Plant/drug effects ; Genes, Reporter ; Glucuronidase/metabolism ; Oryza/enzymology ; Oryza/metabolism ; Plant Proteins/metabolism
    • Abstract:
      Background: Chemically inducible systems that provide both spatial and temporal control of gene expression are essential tools, with many applications in plant biology, yet they have not been extensively tested in monocotyledonous species.
      Results: Using Golden Gate modular cloning, we have created a monocot-optimized dexamethasone (DEX)-inducible pOp6/LhGR system and tested its efficacy in rice using the reporter enzyme β-glucuronidase (GUS). The system is tightly regulated and highly sensitive to DEX application, with 6 h of induction sufficient to induce high levels of GUS activity in transgenic callus. In seedlings, GUS activity was detectable in the root after in vitro application of just 0.01 μM DEX. However, transgenic plants manifested severe developmental perturbations when grown on higher concentrations of DEX. The direct cause of these growth defects is not known, but the rice genome contains sequences with high similarity to the LhGR target sequence lacO, suggesting non-specific activation of endogenous genes by DEX induction. These off-target effects can be minimized by quenching with isopropyl β-D-1-thiogalactopyranoside (IPTG).
      Conclusions: Our results demonstrate that the system is suitable for general use in rice, when the method of DEX application and relevant controls are tailored appropriately for each specific application.
    • References:
      Methods Mol Biol. 2010;655:65-75. (PMID: 20734254)
      Nucleic Acids Res. 2007 Jan;35(Database issue):D883-7. (PMID: 17145706)
      Curr Opin Biotechnol. 2000 Apr;11(2):146-51. (PMID: 10753773)
      PLoS One. 2011 Feb 18;6(2):e16765. (PMID: 21364738)
      Curr Opin Plant Biol. 2003 Apr;6(2):169-77. (PMID: 12667875)
      Plant J. 2006 Feb;45(4):651-83. (PMID: 16441354)
      Planta. 2007 Jul;226(2):453-63. (PMID: 17333253)
      Development. 2006 Oct;133(20):3955-61. (PMID: 16971475)
      Plant J. 1997 Mar;11(3):605-12. (PMID: 9107046)
      PLoS Genet. 2016 Aug 05;12(8):e1006211. (PMID: 27494519)
      Mol Plant Microbe Interact. 2003 Dec;16(12):1069-76. (PMID: 14651340)
      J Exp Bot. 2009;60(6):1799-809. (PMID: 19264754)
      Nucleic Acids Res. 1990 Dec 11;18(23):6767-70. (PMID: 2263444)
      Nucleic Acids Res. 2010 Aug;38(14):e145. (PMID: 20484380)
      Plant J. 2015 May;82(4):707-16. (PMID: 25846505)
      Planta. 2005 Jun;221(4):523-30. (PMID: 15682278)
      Plant J. 2006 Sep;47(6):969-76. (PMID: 16961734)
      Plant J. 2005 Mar;41(6):919-35. (PMID: 15743454)
      PLoS One. 2008;3(11):e3647. (PMID: 18985154)
      ACS Synth Biol. 2016 Jul 15;5(7):632-8. (PMID: 27138234)
      Proc Natl Acad Sci U S A. 1998 Jan 6;95(1):376-81. (PMID: 9419383)
      Front Plant Sci. 2017 Aug 21;8:1447. (PMID: 28871269)
      ACS Synth Biol. 2014 Nov 21;3(11):839-43. (PMID: 24933124)
      EMBO J. 1987 Dec 20;6(13):3901-7. (PMID: 3327686)
      Plant Cell Rep. 2011 Apr;30(4):529-38. (PMID: 21140152)
      Plant J. 1999 Oct;20(1):127-33. (PMID: 10571872)
      Nucleic Acids Res. 2012 Jan;40(Database issue):D1178-86. (PMID: 22110026)
      Nature. 2005 Feb 10;433(7026):629-33. (PMID: 15703747)
      Plant J. 2005 Mar;41(6):899-918. (PMID: 15743453)
      Plant Cell Rep. 2005 May;24(2):86-94. (PMID: 15662501)
      Nucleic Acids Res. 1980 Oct 10;8(19):4321-5. (PMID: 7433111)
      Planta. 2001 Jul;213(3):370-8. (PMID: 11506359)
      Plant Biotechnol J. 2019 Apr 25;:null. (PMID: 31022324)
      Genetics. 2003 Sep;165(1):387-97. (PMID: 14504244)
    • Grant Information:
      OPP 1129902 Bill and Melinda Gates Foundation
    • Contributed Indexing:
      Keywords: Dexamethasone; Golden gate; Oryza sativa, pOp6, LhGR
    • Accession Number:
      0 (Plant Proteins)
      7S5I7G3JQL (Dexamethasone)
      EC 3.2.1.31 (Glucuronidase)
    • Publication Date:
      Date Created: 20191017 Date Completed: 20200127 Latest Revision: 20200127
    • Publication Date:
      20240105
    • Accession Number:
      PMC6794914
    • Accession Number:
      10.1186/s12870-019-2038-x
    • Accession Number:
      31615413