Abstract
BackgroundMouse somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs) by defined factors known to regulate pluripotency, including Oct4, Sox2, Klf4, and c-Myc. It has been reported that Sirtuin 6 (Sirt6), a member of the sirtuin family of NAD+-dependent protein deacetylases, is involved in embryonic stem cell differentiation. However, whether and how Sirt6 influences epigenetic reprogramming remains unknown.MethodsMouse embryonic fibroblasts isolated from transgenic Oct4-GFP reporter mice with or without Sirt6 were used for reprogramming by Yamanaka factors. Alkaline phosphatase-positive and OCT4-GFP-positive colony were counted to calculate reprogramming efficiency. OP9 feeder cell co-culture system was used to measure the hematopoietic differentiation from mouse ES and iPS cells. RNA sequencing was measured to identify the differential expressed genes due to loss of Sirt6 in somatic and pluripotent cells.ResultsIn this study, we provide evidence that Sirt6 is involved in mouse somatic reprogramming. We found that loss of function of Sirt6 could significantly decrease reprogramming efficiency. Furthermore, we showed that Sirt6-null iPS-like cell line has intrinsically a differentiation defect even though the establishment of normal self-renewal. Particularly, by performing transcriptome analysis, we observed that several pluripotent transcriptional factors increase in knockout cell line, which explains the underlying loss of pluripotency in Sirt6-null iPS-like cell line.ConclusionsTaken together, we have identified a new regulatory role of Sirt6 in reprogramming and maintenance of pluripotency.
Highlights
IntroductionMouse somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs) by defined factors known to regulate pluripotency, including Oct, Sox, Klf, and c-Myc. It has been reported that Sirtuin 6 (Sirt6), a member of the sirtuin family of NAD+-dependent protein deacetylases, is involved in embryonic stem cell differentiation
Mouse somatic cells can be reprogrammed into induced pluripotent stem cells by defined factors known to regulate pluripotency, including Oct4, Sox2, Klf4, and c-Myc
The result showed that mRNA level of Sirtuin 6 (Sirt6) was higher expressed by nearly fourfold in ES cell compared to mouse embryonic fibroblast (MEF) (Fig. 1a)
Summary
Mouse somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs) by defined factors known to regulate pluripotency, including Oct, Sox, Klf, and c-Myc. It has been reported that Sirtuin 6 (Sirt6), a member of the sirtuin family of NAD+-dependent protein deacetylases, is involved in embryonic stem cell differentiation. Differentiated somatic cells can be reprogrammed into a pluripotent-like state through four defined factors known to regulate pluripotency, including Oct, Sox, Klf, and c-Myc (OSKM) [1, 2]. Sirt was firstly reported to involve in ES cell anti-oxidation and neural progenitor differentiation [11,12,13,14] Later both our group and others reported that Sirt can promote the efficiency of reprogramming and maintain characteristics of iPS cell [15,16,17]. Whether and how other sirtuins, especially nuclear epigenetic regulator Sirt, regulate mouse somatic reprogramming still remains exclusive
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