Reporter gene silencing in targeted mouse mutants is associated with promoter CpG island methylation

Julia V. Kirov, Michael Adkisson, A. J. Nava, Andreana Cipollone, Brandon Willis, Eric K. Engelhard, Kevin C K Lloyd, Pieter De Jong, David B. West

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Targeted mutations in mouse disrupt local chromatin structure and may lead to unanticipated local effects. We evaluated targeted gene promoter silencing in a group of six mutants carrying the tm1a Knockout Mouse Project allele containing both a LacZ reporter gene driven by the native promoter and a neo selection cassette. Messenger RNA levels of the reporter gene and targeted gene were assessed by qRT-PCR, and methylation of the promoter CpG islands and LacZ coding sequence were evaluated by sequencing of bisulfitetreated DNA. Mutants were stratified by LacZ staining into presumed Silenced and Expressed reporter genes. Silenced mutants had reduced relative quantities LacZ mRNA and greater CpG Island methylation compared with the Expressed mutant group. Within the silenced group, LacZ coding sequence methylation was significantly and positively correlated with CpG Island methylation, while promoter CpG methylation was only weakly correlated with LacZ gene mRNA. The results support the conclusion that there is promoter silencing in a subset of mutants carrying the tm1a allele. The features of targeted genes which promote local silencing when targeted remain unknown.

Original languageEnglish (US)
Article numbere0134155
JournalPLoS One
Volume10
Issue number8
DOIs
StatePublished - Aug 14 2015

Fingerprint

CpG Islands
Methylation
Gene Silencing
gene silencing
Reporter Genes
reporter genes
methylation
Genes
promoter regions
mutants
mice
Lac Operon
Messenger RNA
genes
Alleles
alleles
messenger RNA
DNA Sequence Analysis
Knockout Mice
Chromatin

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Kirov, J. V., Adkisson, M., Nava, A. J., Cipollone, A., Willis, B., Engelhard, E. K., ... West, D. B. (2015). Reporter gene silencing in targeted mouse mutants is associated with promoter CpG island methylation. PLoS One, 10(8), [e0134155]. https://doi.org/10.1371/journal.pone.0134155

Reporter gene silencing in targeted mouse mutants is associated with promoter CpG island methylation. / Kirov, Julia V.; Adkisson, Michael; Nava, A. J.; Cipollone, Andreana; Willis, Brandon; Engelhard, Eric K.; Lloyd, Kevin C K; De Jong, Pieter; West, David B.

In: PLoS One, Vol. 10, No. 8, e0134155, 14.08.2015.

Research output: Contribution to journalArticle

Kirov, JV, Adkisson, M, Nava, AJ, Cipollone, A, Willis, B, Engelhard, EK, Lloyd, KCK, De Jong, P & West, DB 2015, 'Reporter gene silencing in targeted mouse mutants is associated with promoter CpG island methylation', PLoS One, vol. 10, no. 8, e0134155. https://doi.org/10.1371/journal.pone.0134155
Kirov JV, Adkisson M, Nava AJ, Cipollone A, Willis B, Engelhard EK et al. Reporter gene silencing in targeted mouse mutants is associated with promoter CpG island methylation. PLoS One. 2015 Aug 14;10(8). e0134155. https://doi.org/10.1371/journal.pone.0134155
Kirov, Julia V. ; Adkisson, Michael ; Nava, A. J. ; Cipollone, Andreana ; Willis, Brandon ; Engelhard, Eric K. ; Lloyd, Kevin C K ; De Jong, Pieter ; West, David B. / Reporter gene silencing in targeted mouse mutants is associated with promoter CpG island methylation. In: PLoS One. 2015 ; Vol. 10, No. 8.
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