Biotechnology Letters

, Volume 30, Issue 5, pp 885–890 | Cite as

siRNA as a molecular tool for use in Aspergillus niger

  • Sally E. Barnes
  • Marcos J. C. Alcocer
  • David B. Archer
Original Research Paper

Abstract

Gene silencing using siRNA has been examined in the industrially-important fungus, Aspergillus niger. Protoplasts of an A. niger strain containing a single genomic copy of the Escherichia coli uidA gene, encoding β-glucuronidase (GUS), under control of the A. niger glaA promoter at the same genomic locus, were exposed to siRNA targeted against the uidA gene. Down-regulation of uidA mRNA and GUS activity by siRNA was observed in mycelia that developed from the protoplasts. The down-regulation was transient and was not carried over to conidiation. We concluded that gene silencing by siRNA provides a relatively quick method for analysis of gene function in A. niger.

Keywords

Aspergillus niger Down-regulation GUS Protoplasts siRNA 

Notes

Acknowledgements

We acknowledge the financial support of the Biotechnology and Biological Sciences Research Council through the Exploiting Genomics Programme.

References

  1. Archer DB (2000) Filamentous fungi as microbial cell factories for food use. Curr Opin Biotechnol 11:478–483PubMedCrossRefGoogle Scholar
  2. Archer DB, Roberts IN, MacKenzie DA (1990) Heterologous protein secretion from Aspergillus niger in phosphate-buffered batch culture. Appl Microbiol Biotechnol 34:313–315CrossRefGoogle Scholar
  3. Bagasra O, Prilliman KR (2004) RNA interference: the molecular immune system. J Mol Hist 35:545–553CrossRefGoogle Scholar
  4. Catalanotto C, Pallotta M, ReFalo P, Sachs MS, Vayssie L, Macino G, Cogoni C (2004) Redundancy of the two Dicer genes in transgene-induced posttranscriptional gene silencing in Neurospora crassa. Mol Cell Biol 24:2536–2545PubMedCrossRefGoogle Scholar
  5. Chi JT, Chang HY, Wang NN, Chang DS, Dunphy N, Brown PO (2003) Genomewide view of gene silencing by small interfering RNAs. Proc Natl Acad Sci USA 100:6343–6346PubMedCrossRefGoogle Scholar
  6. Cogoni C, Macino G (1999) Gene silencing in Neurospora crassa requires a protein homologous to RNA-dependent RNA polymerase. Nature 399:166–169PubMedCrossRefGoogle Scholar
  7. Cottrell TR, Doering TL (2003) Silence of the strands: RNA interference in eukaryotic pathogens. Trends Microbiol 11:37–43PubMedCrossRefGoogle Scholar
  8. Cove DJ (1966) The induction and repression of nitrate reductase in the fungus Aspergillus nidulans. Biochim Biophys Acta 113:51–56PubMedGoogle Scholar
  9. Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411:494–498PubMedCrossRefGoogle Scholar
  10. Fowler T, Berka RM, Ward M (1990) Regulation of the glaA gene of Aspergillus niger. Curr Genet 18:537–545PubMedCrossRefGoogle Scholar
  11. Hammond SM, Caudy AA, Hannon GJ (2001) Post-transcriptional gene silencing by double-stranded RNA. Nat Rev Genet 2:110–119PubMedCrossRefGoogle Scholar
  12. Hutvagner G, Zamore PD (2002) RNAi: nature abhors a double-strand. Curr Opin Genet Dev 12:225–232PubMedCrossRefGoogle Scholar
  13. Kadotani N, Nakayashiki H, Tosa Y, Shigeyuki M (2003) RNA silencing in the phytopathogenic fungus Magnaporthe oryzae. Mol Plant Microbe Interact 16:769–776PubMedCrossRefGoogle Scholar
  14. Kariko K, Bhuyan P, Capodici J, Ni HP, Lubinski J, Friedman H, Weissman D (2004) Exogenous siRNA mediates sequence-independent gene suppression by signaling through Toll-like receptor 3. Cells Tissues Organs 117:132–138Google Scholar
  15. Liang ZC (2005) High-throughput screening using genome-wide siRNA libraries. IDrugs 8:924–926PubMedGoogle Scholar
  16. Marques JT, Williams BRG (2005) Activation of the mammalian immune system by siRNAs. Nat Biotechnol 23:1399–1405PubMedCrossRefGoogle Scholar
  17. Mouyna I, Henry C, Doering TL, Latge JP (2004) Gene silencing with RNA interference in the human pathogenic fungus Aspergillus fumigatus. FEBS Microbiol Lett 237:317–324Google Scholar
  18. Nakayashiki H (2005) RNA silencing in fungi: Mechanisms and applications. FEBS Lett 579:5950–5957PubMedCrossRefGoogle Scholar
  19. Pel HJ, de Winde JH, Archer DB et al (2007) Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88. Nat Biotechnol 25:221–231PubMedCrossRefGoogle Scholar
  20. Qi YJ, Zhong XH, Itaya A, Ding B (2004) Dissecting RNA silencing in protoplasts uncovers novel effects of viral suppressors on the silencing pathway at the cellular level. Nucleic Acids Res 32:e179PubMedCrossRefGoogle Scholar
  21. Schlaman HR, Risseeuw E, Franke-van Dijk ME, Hooykaas PJ (1994) Nucleotide sequence corrections of the uidA open reading frame encoding β-glucuronidase. Gene 138:259–260PubMedCrossRefGoogle Scholar
  22. Tavernarakis N, Wang SL, Dorovkov M, Ryazanov A, Driscoll M (2000) Heritable and inducible genetic interference by double-stranded RNA encoded by transgenes. Nat Genet 24:180–183PubMedCrossRefGoogle Scholar
  23. Verdoes JC, Punt PJ, Stouthamer AH, van den Hondel CA (1994) The effect of multiple copies of the upstream region on expression of the Aspergillus niger glucoamylase-encoding gene. Gene 145:179–187PubMedCrossRefGoogle Scholar
  24. Waterhouse PM, Graham HW, Wang MB (1998) Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA. Proc Natl Acad Sci USA 98:13959–13964CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Sally E. Barnes
    • 1
  • Marcos J. C. Alcocer
    • 2
  • David B. Archer
    • 1
  1. 1.School of BiologyUniversity of NottinghamNottinghamUK
  2. 2.School of BiosciencesUniversity of NottinghamLoughboroughUK

Personalised recommendations