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Alteration of SC35 localization by transfection reagents

Abstract : Transfection is a powerful tool that enables introducing foreign nucleic acids into living cells in order to study the function of a gene product. Ever since the discovery of transfection many side effects or artifacts caused by transfection reagents have been reported. In this report, we show that the transfection reagent, JetPRIME alters the localization of the splicing protein SC35 widely used as a nuclear speckle marker. We demonstrate that transfection of plasmids with JetPRIME leads to enlarged SC35 speckles and SC35 cytoplasmic granules. By contrast, transfection of the same plasmid with Lipofectamine 3000 does not have any effect on SC35 localization. The formation of SC35 cytoplasmic granules by JetPRIME-mediated transfection is independent of exogenous expression by plasmid and although similar in morphology they are distinct from P-bodies and stress granules. This method of transfection affected only SC35 and phosphorylated SR proteins but not the nuclear speckles. The JetPRIME-mediated transfection also showed compromised transcription in cells with enlarged SC35 speckles. Our work indicates that the use of JetPRIME alters SC35 localization and can affect gene expression and alternative splicing. Therefore, caution should be exercised when interpreting results after the use of a transient transfection system, particularly when the subject of the study is the function of a protein in the control of gene expression or mRNA splicing.
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Submitted on : Tuesday, March 31, 2020 - 11:46:05 AM
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Damodaran et al-2020-Alteratio...
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A.P. Damodaran, T. Courthéoux, E. Watrin, Claude Prigent. Alteration of SC35 localization by transfection reagents. Biochimica et Biophysica Acta - Molecular Cell Research, Elsevier, 2020, 1867 (4), pp.118650. ⟨10.1016/j.bbamcr.2020.118650⟩. ⟨hal-02500548⟩



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