Creation of a novel CRISPR-generated allele to express HA epitope-tagged C1QL1 and improved methods for its detection at synapses

创建一种新的 CRISPR 等位基因来表达 HA 表位标记的 C1QL1,并改进在突触处检测它的方法

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作者: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

Abstract

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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