【系列2】胎牛血清RNA干擾了細(xì)胞培養(yǎng)外源性RNA

FBS-derived miR-1246 is detected in cultured mouse cells

The additional question of broader importance is whether FBS-associated RNA may interfere with cellular RNA analysis. MiR-1246 gene that encodes one of the most abundantly expressed miRNAs in FBS, is present in only 4 out of 223 species, including bovine, human, orangutan and chimpanzee, but not in mouse or rat17. There are no sequences homologous to hsa-miR-1246 identified in the mouse genome. However, low levels of mature miR-1246 were consistently and reproducibly detected in all tested mouse cell lines cultured with 10% FBS (Fig. 1h) by both LNA-based SYBR Green assay (Exiqon) and TaqMan assay (Thermo Fisher Scientific). miR-1246 signal varied between the recipient cells lines, suggesting different levels of uptake and/or processing in the different cells. The miR-1246 signal was significantly reduced in cells that were switched to 10% vdFBS culture medium seven days prior to RNA isolation and undetectable in mouse tissues (Fig. 1h). This example suggests that FBS RNA complexes associated with EVs and possibly non-vesicular RNPs as well, might be taken up by cultured cells and interfere with the quantification of cellular RNA. Considering highly sensitive expression profiling technologies commonly utilized in current research that are capable of detecting femtomoles of RNA, further in-depth analysis of FBS RNA-associated confounders is warranted. As of today, there are no data suggesting the functional activity of this low-level bovine RNA in the recipient cells; nevertheless, subsequent work will be required to accurately address its impact.

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