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Merck
  • Recapitulation of endogenous 4R tau expression and formation of insoluble tau in directly reprogrammed human neurons.

Recapitulation of endogenous 4R tau expression and formation of insoluble tau in directly reprogrammed human neurons.

Cell stem cell (2022-06-07)
Lucia S Capano, Chihiro Sato, Elena Ficulle, Anan Yu, Kanta Horie, Ji-Sun Kwon, Kyle F Burbach, Nicolas R Barthélemy, Susan G Fox, Celeste M Karch, Randall J Bateman, Henry Houlden, Richard I Morimoto, David M Holtzman, Karen E Duff, Andrew S Yoo
摘要

Tau is a microtubule-binding protein expressed in neurons, and the equal ratios between 4-repeat (4R) and 3-repeat (3R) isoforms are maintained in normal adult brain function. Dysregulation of 3R:4R ratio causes tauopathy, and human neurons that recapitulate tau isoforms in health and disease will provide a platform for elucidating pathogenic processes involving tau pathology. We carried out extensive characterizations of tau isoforms expressed in human neurons derived by microRNA-induced neuronal reprogramming of adult fibroblasts. Transcript and protein analyses showed that miR neurons expressed all six isoforms with the 3R:4R isoform ratio equivalent to that detected in human adult brains. Also, miR neurons derived from familial tauopathy patients with a 3R:4R ratio altering mutation showed increased 4R tau and the formation of insoluble tau with seeding activity. Our results collectively demonstrate the utility of miRNA-induced neuronal reprogramming to recapitulate endogenous tau regulation comparable with the adult brain in health and disease.

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Sigma-Aldrich
层粘连蛋白 来源于 Engelbreth-Holm-Swarm 小鼠肉瘤基底膜, 1-2 mg/mL in Tris-buffered saline, 0.2 μm filtered, BioReagent, suitable for cell culture
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N6,2′-O-二丁酰基腺苷 3′,5′-环单磷酸 钠盐, ≥96% (HPLC), powder
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cOmplete Mini蛋白酶抑制剂Cocktail, Tablets provided in a glass vial
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DAPT, ≥98% (HPLC), solid
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纤连蛋白牛血浆, powder, BioReagent, suitable for cell culture
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抗微管相关蛋白2(MAP2)抗体, Chemicon®, from rabbit
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丙戊酸钠盐, A cell-permeable, short-chained fatty acid that inhibits histone deacetylase (IC₅₀ = 400 µM for HDAC1).