Hemagglutinating Virus of Japan Envelope (HVJ-E)-Guided Gene Transfer to the Intestinal Epithelium
Hemagglutinating Virus of Japan Envelope (HVJ-E)-Guided Gene Transfer to the Intestinal Epithelium
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日本包膜血凝病毒 (HVJ-E) 引导基因转移至肠上皮
DOI：
10.1007/978-1-0716-0747-3_19
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Masamichi Imajo
中科院分区：
文献类型：
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作者：
NAKURA MAKOTO;MIYASHITA TOMOHARU;YAMAMOTO YASUHIKO;TAKADA SATOSHI;KANOU SHUNSUKE;TAJIMA HIDEHIRO;TAKAMURA HIROYUKI;OHTA TETSUO;Masamichi Imajo
快速自我更新的小肠上皮为干细胞驱动的组织更新和肿瘤发生提供了一个精致的模型。肠道干细胞(ISCs)位于隐窝的底部，在那里它们产生快速分裂的前体细胞，在几轮细胞分裂时经历细胞周期停滞和终末分化。到目前为止，小鼠的遗传学研究在分析干细胞驱动的肠上皮更新过程中基因的功能方面发挥了核心作用。然而，基因工程小鼠的产生和维护是一项耗时的工作，限制了肠道生物学的进展。最近，我们建立了一种新的方法，作为肠道生物学中小鼠遗传学的替代方法。这种被称为肠特异性基因转移(IGT)的方法，能够利用日本包膜血凝病毒(HVJ-E)将小分子，如siRNAs和质粒，快速有效地运送到活着的小鼠的肠道上皮细胞中。在这里，我们描述了一种详细的IGT方案，并讨论了这种方法如何加速肠道生物学的进展，并阐明肠上皮自我更新的机制。
The rapidly self-renewing epithelium of the small intestine represents an exquisite model for the stem cell-driven tissue renewal and tumorigenesis. Intestinal stem cells (ISCs) are located in the crypt base, where they produce rapidly dividing progenitors that undergo cell-cycle arrest and terminal differentiation upon several rounds of cell division. So far, genetic studies in mice have played a central role in analyzing function of genes during the stem cell-driven renewal of the intestinal epithelium. However, generation and maintenance of genetically engineered mice are a time-consuming endeavor, which limits the progress in intestinal biology. Recently, we have established a novel method that serves as an alternative to mouse genetics in intestinal biology. The method, termed intestine-specific gene transfer (iGT), enables rapid and efficient delivery of small molecules, such as siRNAs and plasmids, into the intestinal epithelium of living mice by utilizing the hemagglutinating virus of Japan envelope (HVJ-E). Here, we describe a detailed protocol for iGT and discuss how this method can accelerate progress in intestinal biology and elucidate the mechanisms of intestinal epithelium self-renewal.