Efficient genetic engineering of murine cochlear organoids

IF 3.1 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of Tissue Engineering and Regenerative Medicine Pub Date : 2022-03-19 DOI:10.1002/term.3298

Yan Zhang, Xiu-Li Hao, Shi-Fang Jia, Yan-Zhen Wen, Ying-Hui Wang

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引用次数: 1

Abstract

Organoid culture is a popular model to study gene function as the easy manipulating and time saving compared with in vivo experiments. This is widely used in auditory system for studying supporting cells (SCs) or hair cells (HCs) as only very few SCs or HCs can be harvested in both human and murine cochlea. However, the use of organoids is still a challenge due to the low efficiency in genetic modification. Here we took Lin28b as an example and compared Lin28b gain-of-function (GOF) and loss-of-function (LOF) with different genetic engineering methods and found that TetOn induced GOF or LOF was more efficient compared with lipofection or lentiviral transduction in the experimental conditions we used. Cell apoptosis in TetOn induction system was lowest compared with the other methods in this study. Our study is the first to compare the efficiency of different genetic engineering techniques in cochlear organoid culture, which may also apply to organoids established with other tissues.

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小鼠耳蜗类器官的高效基因工程研究

与体内实验相比，类器官培养具有操作简便、节省时间等优点，是研究基因功能的一种常用模式。由于在人和鼠耳蜗中只能获得很少的支持细胞或毛细胞，因此这种方法被广泛用于研究听觉系统中的支持细胞或毛细胞。然而，由于基因改造的低效率，类器官的使用仍然是一个挑战。我们以Lin28b为例，比较了不同基因工程方法对Lin28b功能获得(GOF)和功能丧失(LOF)的影响，发现在我们使用的实验条件下，TetOn诱导的GOF或LOF比脂质转染或慢病毒转导更有效。与本研究中其他方法相比，TetOn诱导系统的细胞凋亡最低。我们的研究首次比较了不同基因工程技术在耳蜗类器官培养中的效率，这也可能适用于与其他组织建立的类器官。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Tissue Engineering and Regenerative Medicine 生物-工程：生物医学

CiteScore

7.50

自引率

3.00%

发文量

审稿时长

4-8 weeks

期刊介绍： Journal of Tissue Engineering and Regenerative Medicine publishes rapidly and rigorously peer-reviewed research papers, reviews, clinical case reports, perspectives, and short communications on topics relevant to the development of therapeutic approaches which combine stem or progenitor cells, biomaterials and scaffolds, growth factors and other bioactive agents, and their respective constructs. All papers should deal with research that has a direct or potential impact on the development of novel clinical approaches for the regeneration or repair of tissues and organs. The journal is multidisciplinary, covering the combination of the principles of life sciences and engineering in efforts to advance medicine and clinical strategies. The journal focuses on the use of cells, materials, and biochemical/mechanical factors in the development of biological functional substitutes that restore, maintain, or improve tissue or organ function. The journal publishes research on any tissue or organ and covers all key aspects of the field, including the development of new biomaterials and processing of scaffolds; the use of different types of cells (mainly stem and progenitor cells) and their culture in specific bioreactors; studies in relevant animal models; and clinical trials in human patients performed under strict regulatory and ethical frameworks. Manuscripts describing the use of advanced methods for the characterization of engineered tissues are also of special interest to the journal readership.