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RNAi-Mediated Silencing of FOXO Factors

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1890))

Abstract

RNAi-mediated silencing of a particular gene is an important tool in the field of biology. Knocking down experiments has revealed the role of different proteins and their downstream targets. Here, we describe the procedure of silencing transcription factor of Forkhead box class “O” (FOXO) using shRNA.

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References

  1. Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391(6669):806–811

    Article  CAS  Google Scholar 

  2. Kutter C, Svoboda P (2008) miRNA, siRNA, piRNA: Knowns of the unknown. RNA Biol 5(4):181–188

    Article  CAS  Google Scholar 

  3. Galili N, Davis RJ, Fredericks WJ, Mukhopadhyay S, Rauscher FJ 3rd, Emanuel BS et al (1993) Fusion of a fork head domain gene to PAX3 in the solid tumour alveolar rhabdomyosarcoma. Nat Genet 5(3):230–235

    Article  CAS  Google Scholar 

  4. Hillion J, Le Coniat M, Jonveaux P, Berger R, Bernard OA (1997) AF6q21, a novel partner of the MLL gene in t(6;11)(q21;q23), defines a forkhead transcriptional factor subfamily. Blood 90(9):3714–3719

    Article  CAS  Google Scholar 

  5. Borkhardt A, Repp R, Haas OA, Leis T, Harbott J, Kreuder J et al (1997) Cloning and characterization of AFX, the gene that fuses to MLL in acute leukemias with a t(X;11)(q13;q23). Oncogene 14(2):195–202

    Article  CAS  Google Scholar 

  6. Jacobs FM, van der Heide LP, Wijchers PJ, Burbach JP, Hoekman MF, Smidt MP (2003) FoxO6, a novel member of the FoxO class of transcription factors with distinct shuttling dynamics. J Biol Chem 278(38):35959–35967

    Article  CAS  Google Scholar 

  7. Schmidt M, Fernandez de Mattos S, van der Horst A, Klompmaker R, Kops GJ, Lam EW et al (2002) Cell cycle inhibition by FoxO forkhead transcription factors involves downregulation of cyclin D. Mol Cell Biol 22(22):7842–7852

    Article  CAS  Google Scholar 

  8. Tran H, Brunet A, Grenier JM, Datta SR, Fornace AJ Jr, DiStefano PS et al (2002) DNA repair pathway stimulated by the forkhead transcription factor FOXO3a through the Gadd45 protein. Science 296(5567):530–534

    Article  CAS  Google Scholar 

  9. Zhang X, Tang N, Hadden TJ, Rishi AK (2011) Akt, FoxO and regulation of apoptosis. Biochim Biophys Acta 1813(11):1978–1986

    Article  CAS  Google Scholar 

  10. Gross DN, van den Heuvel AP, Birnbaum MJ (2008) The role of FoxO in the regulation of metabolism. Oncogene 27(16):2320–2336

    Article  CAS  Google Scholar 

  11. Sengupta A, Molkentin JD, Yutzey KE (2009) FoxO transcription factors promote autophagy in cardiomyocytes. J Biol Chem 284(41):28319–28331

    Article  CAS  Google Scholar 

  12. Akhter R, Sanphui P, Biswas SC (2014) The essential role of p53-up-regulated modulator of apoptosis (Puma) and its regulation by FoxO3a transcription factor in beta-amyloid-induced neuron death. J Biol Chem 289(15):10812–10822

    Article  CAS  Google Scholar 

  13. Sanphui P, Biswas SC (2013) FoxO3a is activated and executes neuron death via Bim in response to beta-amyloid. Cell Death Dis 4:e625

    Article  CAS  Google Scholar 

  14. Kyoung Kim H, Kyoung Kim Y, Song IH, Baek SH, Lee SR, Hye Kim J et al (2005) Down-regulation of a forkhead transcription factor, FOXO3a, accelerates cellular senescence in human dermal fibroblasts. J Gerontol A Biol Sci Med Sci 60(1):4–9

    Article  Google Scholar 

  15. Ui-Tei K, Naito Y, Takahashi F, Haraguchi T, Ohki-Hamazaki H, Juni A et al (2004) Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference. Nucleic Acids Res 32(3):936–948

    Article  CAS  Google Scholar 

  16. Taxman DJ, Livingstone LR, Zhang J, Conti BJ, Iocca HA, Williams KL et al (2006) Criteria for effective design, construction, and gene knockdown by shRNA vectors. BMC Biotechnol 6:7

    Article  Google Scholar 

  17. Reynolds A, Leake D, Boese Q, Scaringe S, Marshall WS, Khvorova A (2004) Rational siRNA design for RNA interference. Nat Biotechnol 22(3):326–330

    Article  CAS  Google Scholar 

  18. Biswas SC, Shi Y, Sproul A, Greene LA (2007) Pro-apoptotic Bim induction in response to nerve growth factor deprivation requires simultaneous activation of three different death signaling pathways. J Biol Chem 282(40):29368–29374

    Article  CAS  Google Scholar 

  19. Amarzguioui M, Prydz H (2004) An algorithm for selection of functional siRNA sequences. Biochem Biophys Res Commun 316(4):1050–1058

    Article  CAS  Google Scholar 

  20. Hribal ML, Nakae J, Kitamura T, Shutter JR, Accili D (2003) Regulation of insulin-like growth factor-dependent myoblast differentiation by Foxo forkhead transcription factors. J Cell Biol 162(4):535–541

    Article  CAS  Google Scholar 

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Acknowledgement

The work was supported by one of the 12th Five Year Plan Projects, miND (BSC0115) of CSIR, Govt. of India.

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Correspondence to Subhas Chandra Biswas .

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Sanphui, P., Saha, A., Biswas, S.C. (2019). RNAi-Mediated Silencing of FOXO Factors. In: Link, W. (eds) FOXO Transcription Factors. Methods in Molecular Biology, vol 1890. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8900-3_11

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  • DOI: https://doi.org/10.1007/978-1-4939-8900-3_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-8899-0

  • Online ISBN: 978-1-4939-8900-3

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