Activation tag screening for cell expansion genes in Arabidopsis thaliana

Chaowen Xiao, Charles T. Anderson

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Forward genetic screens for growth-deficient loss-of-function mutants have uncovered a wide array of genes involved in cell expansion. However, the centrality of cell growth to plant survival means that null mutations in many genes involved in this process are likely to be lethal early in development, making phenotypic analysis difficult. Additionally, the phenotypes of loss-of-function mutations in genes that are members of large gene families might be masked by functional redundancy with other family members. Activation tagging provides a method of screening for dominant overexpression phenotypes in an arbitrarily large collection of transgenic individuals, allowing for functional genomic identification of genes related to cell growth and expansion. In this chapter, we discuss the advantages and limitations of activation tag screening and describe a protocol for identifying activation tag lines with enhanced cell expansion, using dark-grown Arabidopsis thaliana seedlings as an experimental system. We also describe secondary screens to identify candidate genes for further cell biological and genetic characterization. These protocols can be adapted to any process or species of interest, as long as a suitable activation-tagged population and a genome sequence are available.

Original languageEnglish (US)
Pages (from-to)159-171
Number of pages13
JournalMethods in Molecular Biology
Volume1242
DOIs
StatePublished - Jan 1 2015

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Arabidopsis
Genes
Growth
Phenotype
Mutation
Seedlings
Genome
Population

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

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abstract = "Forward genetic screens for growth-deficient loss-of-function mutants have uncovered a wide array of genes involved in cell expansion. However, the centrality of cell growth to plant survival means that null mutations in many genes involved in this process are likely to be lethal early in development, making phenotypic analysis difficult. Additionally, the phenotypes of loss-of-function mutations in genes that are members of large gene families might be masked by functional redundancy with other family members. Activation tagging provides a method of screening for dominant overexpression phenotypes in an arbitrarily large collection of transgenic individuals, allowing for functional genomic identification of genes related to cell growth and expansion. In this chapter, we discuss the advantages and limitations of activation tag screening and describe a protocol for identifying activation tag lines with enhanced cell expansion, using dark-grown Arabidopsis thaliana seedlings as an experimental system. We also describe secondary screens to identify candidate genes for further cell biological and genetic characterization. These protocols can be adapted to any process or species of interest, as long as a suitable activation-tagged population and a genome sequence are available.",
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Activation tag screening for cell expansion genes in Arabidopsis thaliana. / Xiao, Chaowen; Anderson, Charles T.

In: Methods in Molecular Biology, Vol. 1242, 01.01.2015, p. 159-171.

Research output: Contribution to journalArticle

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