1. Generation of Stable Cell Lines of APP695 (K595N, M596L) Gene Over-Expression via Lentiviral Over-Expression System.
- Author
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Huitao Zhang, Hao, Rui, Yu, Zitong, Hu, Shi, Ren, Jingyi, Hong, Yanhang, Wang, Shifan, Sun, Huihui, Zhang, Pengcheng, Zhang, Yi, Yang, Hui, and Ding, Juan
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CELL lines , *CELL culture , *ENZYME-linked immunosorbent assay , *ALZHEIMER'S disease , *PROTEIN precursors , *GENE expression - Abstract
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 Aβ1-42, respectively, in addition, the expression of Aβ1-42 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. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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