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Related Experiment Video

Updated: Sep 29, 2025

Culture of Embryonic Mouse Cochlear Explants and Gene Transfer by Electroporation
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Efficient genetic engineering of murine cochlear organoids.

Yan Zhang1, Xiu-Li Hao1, Shi-Fang Jia1

  • 1School of Chemical and Biological Engineering, Tai Yuan University of Science and Technology, Taiyuan, China.

Journal of Tissue Engineering and Regenerative Medicine
|March 19, 2022
PubMed
Summary

TetOn gene induction is the most efficient method for genetic modification in cochlear organoids, outperforming other techniques and minimizing cell apoptosis. This finding enhances gene function studies in auditory research.

Keywords:
LIN28cochleagenetic engineeringorganoid

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Area of Science:

  • Developmental Biology
  • Genetics
  • Otolaryngology

Background:

  • Organoid cultures offer a viable in vivo alternative for studying gene function, particularly in the auditory system where primary cell isolation is limited.
  • Genetic modification of cochlear organoids is crucial for understanding supporting cells (SCs) and hair cells (HCs) but faces efficiency challenges.

Purpose of the Study:

  • To compare the efficiency of different genetic engineering methods for gene manipulation in cochlear organoids.
  • To evaluate the impact of Lin28b gain-of-function (GOF) and loss-of-function (LOF) using various genetic engineering techniques.

Main Methods:

  • Utilized TetOn-inducible systems, lipofection, and lentiviral transduction for genetic modification.
  • Assessed gene editing efficiency and cellular apoptosis rates across different methods.
  • Focused on Lin28b as a model gene for gain-of-function and loss-of-function studies.

Main Results:

  • TetOn-induced GOF and LOF demonstrated higher efficiency compared to lipofection and lentiviral transduction under the tested conditions.
  • The TetOn induction system exhibited the lowest rate of cell apoptosis among the evaluated methods.
  • This study provides the first comparative analysis of genetic engineering techniques in cochlear organoids.

Conclusions:

  • TetOn-inducible systems represent a superior strategy for efficient genetic engineering in cochlear organoids.
  • The findings offer a valuable approach for advancing gene function studies in auditory organoid models and potentially other tissue-derived organoids.