Advertisement

Studying Autoimmunity by In Vivo RNA Interference

  • Stephan Kissler
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 555)

Abstract

The occurrence of autoimmunity is strongly associated with multiple gene variants that predispose individuals to disease. The identification of the gene polymorphisms that modulate disease susceptibility is key to our understanding of disease etiology and pathogenesis. While genetic studies in humans have uncovered several associations and have provided possible candidate genes for further study, the use of animal models is indispensable for detailed functional studies. In order to facilitate the genetic manipulation of experimental models of autoimmunity, we employ lentiviral transgenesis in combination with RNA interference (RNAi). This approach bypasses the need for targeted mutagenesis of embryonic stem cells and/or backcrossing of genetically modified animals onto the relevant genetic background. Lentiviral RNAi offers several advantages compared to conventional transgenesis or knockout technology, and these, as well as the technique’s weaknesses, are discussed herein.

Key words

Lentivirus RNAi autoimmunity transgenesis mouse embryo 

References

  1. 1.
    Lois C., Hong E.J., Pease S., Brown E.J. and Baltimore D. (2002) Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science 295: 868–872.PubMedCrossRefGoogle Scholar
  2. 2.
    Brummelkamp T.R., Bernards R. and Agami R. (2002) A system for stable expression of short interfering RNAs in mammalian cells. Science 296: 550–553.PubMedCrossRefGoogle Scholar
  3. 3.
    Rubinson et al. (2003) A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Nat. Genet.33: 401–406.PubMedCrossRefGoogle Scholar
  4. 4.
    Tiscornia G., Singer O., Ikawa M. and Verma I.M. (2003) A general method for gene knockdown in mice by using lentiviral vectors expressing small interfering RNA. Proc. Natl. Acad. Sci. USA 100: 1844–1848.PubMedCrossRefGoogle Scholar
  5. 5.
    Ventura A., Meissner A., Dillon C.P., McManus M.T., Sharp P.A., Van Parijs L., Jaenisch R. and Jacks T. (2004) Cre-lox conditional RNA interference from transgenes. Proc. Natl. Acad. Sci. USA 101: 10380–10385.PubMedCrossRefGoogle Scholar
  6. 6.
    Szulc J., Wiznerowicz M., Sauvain M.-O., Trono D., and Aebischer P. (2006) A versatile tool for conditional gene expression and knockdown. Nat. Methods 3: 109–116.PubMedCrossRefGoogle Scholar
  7. 7.
    Kissler S., Stern P., Takahashi K., Hunter K., Peterson L.B. and Wicker L.S. (2006) In vivo RNA interference demonstrates a role for Nramp1 in modifying susceptibility to type 1 diabetes. Nat. Genet.38: 479–483.PubMedCrossRefGoogle Scholar
  8. 8.
    Stegmeier F., Hu G., Rickles R.J., Hannon G.J., and Elledge S.J. (2005) A lentiviral microRNA-based system for single-copy polymerase II-regulated RNA interference in mammalian cells. Proc. Natl. Acad. Sci. USA102: 13212–13217.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Stephan Kissler
    • 1
  1. 1.Rudolf Virchow Center / DFG Center for Experimental BiomedicineUniversity of WürzburgWürzburgGermany

Personalised recommendations