siRNA-mediated gene silencing in the salivary gland using in vivo microbubble-enhanced sonoporation

  1. T Sakai1,2,
  2. M Kawaguchi1,
  3. Y Kosuge3

Article first published online: 10 JUN 2009

DOI: 10.1111/j.1601-0825.2009.01579.x

Issue

Oral Diseases

Oral Diseases

Volume 15, Issue 7, pages 505–511, October 2009

Additional Information

How to Cite

Sakai, T., Kawaguchi, M. and Kosuge, Y. (2009), siRNA-mediated gene silencing in the salivary gland using in vivo microbubble-enhanced sonoporation. Oral Diseases, 15: 505–511. doi: 10.1111/j.1601-0825.2009.01579.x

Author Information

  1. 1

    Department of Pharmacology

  2. 2

    Oral Health Science Center hrc7, Tokyo Dental College, Chiba

  3. 3

    Research Unit of Pharmacology, College of Pharmacy, Nihon University, Funabasi, Chiba, Japan

*T Sakai, Department of Pharmacology, Tokyo Dental College, 1-2-2 Masago, Mihama-ku, Chiba, Chiba, 261-8502 Japan. Tel: +81 43 270 3774, Fax: +81 43 270 3776, E-mail: takasakai@tdc.ac.jp

Publication History

  1. Issue published online: 10 SEP 2009
  2. Article first published online: 10 JUN 2009
  3. Received 15 October 2008; revised 20 February 2009; accepted 28 April 2009

SEARCH

SEARCH BY CITATION

ARTICLE TOOLS

View Full Article (HTML) Get PDF (796K)

Keywords:

  • sonoporation;
  • siRNA;
  • gene silencing;
  • in vivo;
  • salivary gland

Objectives:  siRNA-induced gene silencing in the salivary gland using microbubble-enhanced sonoporation was used to develop an in vivo gene knockdown technique.

Methods:  siRNA targeting rat glyceraldehyde-3-phosphate dehydrogenas (GAPDH) was mixed with echo-enhanced microbubbles and reverse-injected into rat parotid glands using transdermal ultrasound. To compare direct and transdermal ultrasound efficiencies, an incision was made on the lateral neck to expose the parotid glands for direct application. The efficiency of gene suppression was determined using quantitative reverse transcription-polymerase chain reaction 24–72 h after siRNA delivery. Cytotoxicity was assessed using histological analysis.

Results:  Expression of rat GAPDH in the parotid glands was silenced 48 h after siRNA was delivered by ultrasound (frequency: 1 MHz; intensity: 2 W cm−2; exposure time: 2 min). High-intensity ultrasound induced tissue damage and apoptotic change. Echo-enhanced microbubbles significantly improved siRNA-induced gene silencing by 10–50%. Compared with transdermal application, direct-exposure ultrasound was only slightly effective, and no significant difference in gene expression was observed.

Conclusion:  The results indicate that microbubble-enhanced sonoporation can yield in vivo siRNA gene silencing in the rat parotid gland. This technique could be applied to provide gene knockdown organs for functional genomic analyses and to develop siRNA-based gene therapy.

View Full Article (HTML) Get PDF (796K)