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- W2919448418 abstract "Abstract MicroRNAs (miRNAs) bind to the 3ʹ-untranslated region of target mRNAs in a sequence-specific manner and subsequently repress gene translation. Human miR-26a has been studied extensively, but the target transcripts are far from complete. We first employed the CRISPR-Cas9 system to generate an miR-26a-knockout line in human cervical cancer HeLa cells. The miR26a-knockout line showed increased cell growth and altered proliferation. Proteomics technology of sequential window acquisition of all theoretical mass spectra (SWATH-MS) was utilized to compare the protein abundance between the wild-type and the knockout lines, with an attempt to identify transcripts whose translation was influenced by miR-26a. Functional classification of the proteins with significant changes revealed their function in stress response, proliferation, localization, development, signaling, etc. Several proteins in the cell cycle/proliferation signaling pathway were chosen to be validated by western blot and parallel reaction monitoring (PRM). The satisfactory consistency among the three approaches indicated the reliability of the SWATH-MS quantification. Among the computationally predicted targets, a subset of the targets was directly regulated by miR-26a, as demonstrated by luciferase assays and Western blotting. This study creates an inventory of miR-26a-targeted transcripts in HeLa cells and provides fundamental knowledge to further explore the functions of miR-26a in human cancer." @default.
- W2919448418 created "2019-03-11" @default.
- W2919448418 creator A5008463603 @default.
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- W2919448418 date "2019-02-04" @default.
- W2919448418 modified "2023-10-16" @default.
- W2919448418 title "Label-free Quantitative Analysis of Protein Expression Alterations in miR-26a-Knockout HeLa Cells using SWATH-MS Technology" @default.
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- W2919448418 doi "https://doi.org/10.1038/s41598-018-34904-8" @default.
- W2919448418 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6362012" @default.
- W2919448418 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/30718521" @default.
- W2919448418 hasPublicationYear "2019" @default.
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