gms | German Medical Science

83. Jahresversammlung der Deutschen Gesellschaft für Hals-Nasen-Ohren-Heilkunde, Kopf- und Hals-Chirurgie e. V.

Deutsche Gesellschaft für Hals-Nasen-Ohren-Heilkunde, Kopf- und Hals-Chirurgie e. V.

16.05. - 20.05.2012, Mainz

Intratympanic injection of shRNA-expressing lentivirus causes gene silencing in the inner ear in chicken

Meeting Abstract

Suche in Medline nach

  • corresponding author Mingliang Xiang - Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, P.R. China
  • Yu Jia - Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, P.R. China
  • Chenling Shen - Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, P.R. China
  • Hao Wu - Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, P.R. China

Deutsche Gesellschaft für Hals-Nasen-Ohren-Heilkunde, Kopf- und Hals-Chirurgie. 83. Jahresversammlung der Deutschen Gesellschaft für Hals-Nasen-Ohren-Heilkunde, Kopf- und Hals-Chirurgie. Mainz, 16.-20.05.2012. Düsseldorf: German Medical Science GMS Publishing House; 2012. Doc12hnod573

doi: 10.3205/12hnod573, urn:nbn:de:0183-12hnod5739

Veröffentlicht: 4. April 2012

© 2012 Xiang et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielf&aauml;ltigt, verbreitet und &oauml;ffentlich zug&aauml;nglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

Text

Objectives/Hypothesis: To test if intratympanic injection of shRNA-expressing lentivirus can cause gene silencing in the inner ear in chicken.

Study Design: Randomized controlled trial.

Methods: EFNA2 was selected as the target gene for shRNA-mediated silencing. pFU-GW-iRNA viral vector and cationic lipid complex method were used to construct and package the shRNA-expressing lentivirus. The packaged virus was directly injected into the middle ear of chickens at the age of two weeks. Fluorescence imaging was used to determine if the inner cells were transfected by the virus. Quantitative RT-PCR was employed to examine the silencing effect.

Results: In total, three plasmids expressing EFNA2-targeting shRNA were synthesized and tested to be effective in vitro. The one with the highest interfering efficiency (76% reduction) was selected for intratympanic microinjection. Fluorescence imaging showed that the lentivirus infected the inner ear widely. Quantitative RT-PCR demonstrated that the mRNA level of EFNA2 was dramatically reduced by 73%.

Conclusions: shRNA-expressing lentivirus injected into the middle ear through tympanic membrane can spread into and infect the inner ear tissue to cause gene silencing in chicken, therefore we established a convenient way for local manipulation of genes in the inner ear.

Supported by: grant No. 08ZR1414900 from the Science and Technology Commission of Shanghai Municipality, China. The authors have no other funding, financial relationships, or conflicts of interest to disclose.