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Application of lentivirus-mediated RNAi in studying gene function in mammalian tooth development

  1. Yiqiang Song1,§,
  2. Zunyi Zhang1,†,§,
  3. Xueyan Yu1,
  4. Minquan Yan1,
  5. Xiaoyun Zhang1,†,
  6. Shuping Gu1,‡,
  7. Thomas Stuart2,
  8. Chao Liu3,
  9. Jakob Reiser4,
  10. Yanding Zhang3,
  11. Yiping Chen1,3,*

Article first published online: 20 APR 2006

DOI: 10.1002/dvdy.20706

Issue

Developmental Dynamics

Developmental Dynamics

Special Issue: Craniofacial Development Special Issue

Volume 235, Issue 5, pages 1347–1357, May 2006

Additional Information

How to Cite

Song, Y., Zhang, Z., Yu, X., Yan, M., Zhang, X., Gu, S., Stuart, T., Liu, C., Reiser, J., Zhang, Y. and Chen, Y. (2006), Application of lentivirus-mediated RNAi in studying gene function in mammalian tooth development. Dev. Dyn., 235: 1347–1357. doi: 10.1002/dvdy.20706

Author Information

  1. 1

    Division of Developmental Biology, Department of Cell and Molecular Biology, Tulane University, New Orleans, Louisiana

  2. 2

    Molecular Neurobiotechnology Core Facility, Department of Cell and Molecular Biology, Tulane University, New Orleans, Louisiana

  3. 3

    College of Bioengineering, Fujian Normal University, Fuzhou, Fujian, P.R. China

  4. 4

    Gene Therapy Program, Louisiana State University Health Sciences Center, New Orleans, Louisiana

  1. †

    Center for Oral Biology, University of Rochester, School of Medicine and Dentistry, Rochester, NY 14642

  2. ‡

    Shanghai Research Center for Biomodel Organism, 88 Cailun Road, Pudong, Shanghai, China 201203

  1. §

    Yiqiang Song and Zunyi Zhang contributed equally to this work.

*Department of Cell and Molecular Biology, Tulane University, New Orleans, LA 70118

Publication History

  1. Issue published online: 20 APR 2006
  2. Article first published online: 20 APR 2006
  3. Manuscript Accepted: 30 DEC 2005

Funded by

  • National Institutes of Health. Grant Numbers: DE16623, DE15123, DE12329
  • Millennium Trust Health Excellent Fund. Grant Number: HEF-2000-05-04
  • National Natural Science Foundation of China. Grant Numbers: 30270652, 30370705
  • Fujian Provincial Department of Science and Technology, China. Grant Numbers: 2002I006, C0320002

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Keywords:

  • RNAi;
  • lentivirus;
  • gene function;
  • tooth development;
  • Barx1;
  • Dlx;
  • Msx1

Abstract

RNA interference (RNAi) has recently become a powerful tool to silence gene expression in mammalian cells, but its application in assessing gene function in mammalian developing organs remains highly limited. Here we describe several unique developmental properties of the mouse molar germ. Embryonic molar mesenchyme, but not the incisor mesenchyme, once dissociated into single cell suspension and re-aggregated, retains its odontogenic potential, the capability of a tissue to instruct an adjacent tissue to initiate tooth formation. Dissociated molar mesenchymal cells, even after being plated in cell culture, retain odontogenic competence, the capability of a tissue to respond to odontogenic signals and to support tooth formation. Most interestingly, while dissociated epithelial and mesenchymal cells of molar tooth germ are mixed and re-aggregated, the epithelial cells are able to sort out from the mesenchymal cells and organize into a well-defined dental epithelial structure, leading to the formation of a well-differentiated tooth organ after sub-renal culture. These unique molar developmental properties allow us to develop a strategy using a lentivirus-mediated RNAi approach to silence gene expression in dental mesenchymal cells and assess gene function in tooth development. We show that knockdown of Msx1 or Dlx2 expression in the dental mesenchyme faithfully recapitulates the tooth phenotype of their targeted mutant mice. Silencing of Barx1 expression in the dental mesenchyme causes an arrest of tooth development at the bud stage, demonstrating a crucial role for Barx1 in tooth formation. Our studies have established a reliable and rapid assay that would permit large-scale analysis of gene function in mammalian tooth development. Developmental Dynamics 235:1334–1344, 2006. © 2006 Wiley-Liss, Inc.

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