创建 CRISPR 生成的新型等位基因,以表达 HA 表位标记的 C1QL1,并改进其在突触处的检测方法。

IF 3.5 4区 生物学 Q1 Biochemistry, Genetics and Molecular Biology
Hiu W Cheung, Alexander D Schouw, Zeynep M Altunay, J Wesley Maddox, Lyndsay C Kresic, Brenna C McAllister, Keaven Caro, Shahnawaz Alam, Angie Huang, Robert S Pijewski, Amy Lee, David C Martinelli
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引用次数: 0

摘要

C1QL1 在大脑的一部分细胞中表达,可能具有多种功能,包括调节神经元之间的突触。由于缺乏可用的抗体,C1QL 蛋白的研究进展缓慢。因此,我们创建了一种新型基因敲入小鼠品系,在该品系中,内源性 C1ql1 基因座插入了一个 HA 标记。我们对整个大脑进行了检查,发现了以前未被重视的表达 C1QL1 的核团,推测这些核团存在于神经元中。然而,从总数来看,绝大多数表达 C1QL1 的细胞属于少突胶质细胞系。突触裂隙蛋白的亚细胞免疫定位具有挑战性,因此我们开发了一种新的方案来改善突触处的信号。最后,我们对各种抗HA抗体进行了比较,以帮助未来使用这种HA表位标记等位基因进行研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Creation of a novel CRISPR-generated allele to express HA epitope-tagged C1QL1 and improved methods for its detection at synapses.

C1QL1 is expressed in a subset of cells in the brain and likely has pleiotropic functions, including the regulation of neuron-to-neuron synapses. Research progress on C1QL proteins has been slowed by a dearth of available antibodies. Therefore, we created a novel knock-in mouse line in which an HA-tag is inserted into the endogenous C1ql1 locus. We examined the entire brain, identifying previously unappreciated nuclei expressing C1QL1, presumably in neurons. By total numbers, however, the large majority of C1QL1-expressing cells are of the oligodendrocyte lineage. Subcellular immunolocalization of synaptic cleft proteins is challenging, so we developed a new protocol to improve signal at synapses. Lastly, we compared various anti-HA antibodies to assist future investigations using this and likely other HA epitope-tagged alleles.

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来源期刊
FEBS Letters
FEBS Letters 生物-生化与分子生物学
CiteScore
7.00
自引率
2.90%
发文量
303
审稿时长
1.0 months
期刊介绍: FEBS Letters is one of the world''s leading journals in molecular biology and is renowned both for its quality of content and speed of production. Bringing together the most important developments in the molecular biosciences, FEBS Letters provides an international forum for Minireviews, Research Letters and Hypotheses that merit urgent publication.
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