Molecular Biotechnology

, Volume 48, Issue 1, pp 38–48

RNA Interference in Pigs: Comparison of RNAi Test Systems and Expression Vectors

  • Claudia Merkl
  • Simon Leuchs
  • Anja Saalfrank
  • Alexander Kind
  • Angelika Schnieke
Research

DOI: 10.1007/s12033-010-9346-6

Cite this article as:
Merkl, C., Leuchs, S., Saalfrank, A. et al. Mol Biotechnol (2011) 48: 38. doi:10.1007/s12033-010-9346-6

Abstract

We have examined the use of RNA interference as a means of downregulating gene expression and provide the first comparison of shRNA and artificial miRNA constructs for transgenic livestock. Several in vitro assays were performed to identify the most effective RNAi constructs. shRNA and miRNA constructs achieved significant downregulation of two porcine target genes: the milk whey protein beta-lactoglobulin and the tumour suppressor p53. Results of different assays were, however, sometimes at variance, indicating that no one assay can be relied upon to predict the effectiveness of an RNAi construct. Our findings are that screening of RNAi constructs is most informative if carried out in primary cells that express the target gene and are competent for somatic cell nuclear transfer. Importantly, the use of miRNA constructs makes tissue specific gene knockdown in large animals a realistic possibility.

Keywords

RNA interferencemiRNAshRNAPigTransgenic animalsp53Beta-lactoglobulinPredictive dataIn vitro assay

Supplementary material

12033_2010_9346_MOESM1_ESM.doc (75 kb)
Supplementary material 1 (DOC 75 kb)

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Claudia Merkl
    • 1
  • Simon Leuchs
    • 1
  • Anja Saalfrank
    • 1
  • Alexander Kind
    • 1
  • Angelika Schnieke
    • 1
  1. 1.Chair of Livestock Biotechnology, TU MuenchenFreisingGermany