Abstract
RNA interference (RNAi) pathways regulate self-renewal and differentiation of embryonic stem (ES) cells. Argonaute 2 (Ago2) is a vital component of RNA-induced silencing complex (RISC) and the only Ago protein with slicer activity. We generated Ago2-deficient ES cells by conditional gene targeting. Ago2-deficient ES cells are defective in the small-RNA-mediated gene silencing and are significantly compromised in biogenesis of mature microRNA. The self-renewal rate of Ago2-deficient ES cells is affected due to failure of silencing of Cdkn1a by ES-cell-specific microRNAs (miRNA) in the absence of Ago2. Interestingly, unlike Dicer- and Dgcr8-deficient ES cells, they differentiate to all three germ layers both in vivo and in vitro. However, early differentiation of Ago2-deficient ES cells is delayed by 2–4 days as indicated by persistence of higher levels of self-renewal/ pluripotency markers during differentiation. Further, appearance of morphological and differentiation markers is also delayed during the differentiation. In this study we show that Ago2 is essential for normal self-renewal and differentiation. Also, our data suggest that self-renewal and differentiation of ES cells are regulated by both siRNA and miRNA pathways.
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Acknowledgements
We thank David M Gilbert and Jyotsna Dhawan for suggestions and reading the manuscript. We thank Jedy Jose for excellent technical assistance. This work was supported by the Council of Scientific and Industrial Research, India.
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Corresponding editor: Prim B Singh
[Chandra Shekar P, Naim A, Partha Sarathi D and Kumar S 2011 Argonaute-2-null embryonic stem cells are retarded in self-renewal and differentiation. J. Biosci. 36 649–657] DOI 10.1007/s12038-011-9094-1
Supplementary materials pertaining to this article are available on the Journal of Biosciences Website at http://www.ias.ac.in/jbiosci/Sep2011/pp649–657/suppl.pdf
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Shekar, P.C., Naim, A., Sarathi, D.P. et al. Argonaute-2-null embryonic stem cells are retarded in self-renewal and differentiation. J Biosci 36, 649–657 (2011). https://doi.org/10.1007/s12038-011-9094-1
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DOI: https://doi.org/10.1007/s12038-011-9094-1