Transgene-mediated co-suppression of DNA topoisomerase-1 gene in caenorhabditis elegans

Myon Hee Lee, Dong Seok Cha, Srivalli Swathi Mamillapalli, Young Chul Kwon, Hyeon-Sook Koo

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Ectopic expression of multi-transgenic copies can result in reduced expression of the transgene and can induce silence of endogenous gene; this process is called as co-suppression. Using a transgene-mediated co-suppression technique, we demonstrated the biological function of DNA topoisomerase-1 (top-1) in C. elegans development. Introduction of full-length top-1 transgene sufficiently induced the co-suppression of endogenous top-1 gene, causing embryonic lethality and abnormal germline development. We also found that the co-suppression of top-1 gene affected morphogenesis, lifespan and larval growth that were not observed in top-1(RNAi) animals. Strikingly, co-suppression effects were significantly reduced by the elimination of top-1 introns, suggesting that ef-ficient co-suppression may require intron(s) in C. elegans. Sequence analysis revealed that the introns 1 and 2 of top-1 gene possess consensus binding sites for several transcription factors, including MAB-3, LIN-14, TTX-3/CEH-10, CEH-1, and CEH-22. Among them, we examined a genetic link between ceh-22 and top-1. The ceh-22 is partially required for the specification of distal tip cells (DTC), which functions as a stem cell niche in the C. elegans gonad. Intriguingly, top-1(RNAi) significantly enhanced DTC loss in ceh-22 mutant gonads, indicating that top-1 may play an important role in CEH-22-mediated DTC fate specification. Therefore, our findings suggest that transgene-mediated co-suppression facilitates the silencing of the specific genes and the study of gene function in vivo.

Original languageEnglish
Pages (from-to)11-20
Number of pages10
JournalInternational Journal of Biochemistry and Molecular Biology
Volume5
Issue number1
Publication statusPublished - 2014 Jan 1

Fingerprint

Type I DNA Topoisomerase
Caenorhabditis elegans
RNA Interference
Transgenes
Genes
Introns
Gonads
Specifications
Stem cells
Stem Cell Niche
Animals
Transcription Factors
Binding Sites
Gene Silencing
Morphogenesis
Sequence Analysis

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Lee, Myon Hee ; Cha, Dong Seok ; Mamillapalli, Srivalli Swathi ; Kwon, Young Chul ; Koo, Hyeon-Sook. / Transgene-mediated co-suppression of DNA topoisomerase-1 gene in caenorhabditis elegans. In: International Journal of Biochemistry and Molecular Biology. 2014 ; Vol. 5, No. 1. pp. 11-20.
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Transgene-mediated co-suppression of DNA topoisomerase-1 gene in caenorhabditis elegans. / Lee, Myon Hee; Cha, Dong Seok; Mamillapalli, Srivalli Swathi; Kwon, Young Chul; Koo, Hyeon-Sook.

In: International Journal of Biochemistry and Molecular Biology, Vol. 5, No. 1, 01.01.2014, p. 11-20.

Research output: Contribution to journalArticle

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