Generation of Stable Cell Lines of APP695 (K595N, M596L) Gene Over-Expression via Lentiviral Over-Expression System

There are now various theories on the pathophysiology of Alzheimer’s disease (AD), the β-Amyloid (Aβ) cascade hypothesis has garnered a lot of interest. This text applied lentivirus-encapsulated plasmid to transfect APP695 (K595N, M596L) gene into SH-SY5Y cells and thereby obtain a stable cell line...

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Bibliographic Details
Published in:Biology bulletin of the Russian Academy of Sciences 2024, Vol.51 (1), p.30-36
Main Authors: Huitao Zhang, Hao, Rui, Yu, Zitong, Hu, Shi, Ren, Jingyi, Hong, Yanhang, Wang, Shifan, Sun, Huihui, Zhang, Pengcheng, Zhang, Yi, Yang, Hui, Ding, Juan
Format: Article
Language:English
Subjects:
Alzheimer's disease
Amyloid precursor protein
Biochemistry
Biomedical and Life Sciences
Cell Biology
Cell culture
Cell lines
Ecology
Enzyme-linked immunosorbent assay
Gene expression
Life Sciences
Neurodegenerative diseases
Overexpression
Transfection
Zoology
β-Amyloid
β-Site APP-cleaving enzyme 1
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Summary:There are now various theories on the pathophysiology of Alzheimer’s disease (AD), the β-Amyloid (Aβ) cascade hypothesis has garnered a lot of interest. This text applied lentivirus-encapsulated plasmid to transfect APP695 (K595N, M596L) gene into SH-SY5Y cells and thereby obtain a stable cell line over-expressing Aβ. We used qPCR to monitor the mRNA expression of β-Amyloid Precursor Protein (APP) gene, β-Site APP Cleaving Enzyme 1 (BACE1) gene, we also used Western Blot to detect the APP protein, BACE1 protein, and Aβ 1-42 , respectively, in addition, the expression of Aβ 1-42 in SH-SY5Y cell lysate and cell culture supernatant was measured by Enzyme-Linked Immunosorbent Assay (ELISA). The findings demonstrated APP gene over-expression following APP695 (K595N, M596L) gene transfection, and also indicated the successful establishment of the Swedish mutant cell model.
ISSN:1062-3590
1608-3059
DOI:10.1134/S1062359023602586