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Volume 37 Issue 3
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Article Navigation >  Journal of Pharmaceutical Practice and Service  > 2019  >  37(3): 237-240

WANG Zhi, WANG Shuna, XU Tianying, LI Zhiyong, ZHANG Sailong, MIAO Chaoyu. The effect of Nampt gene editing on the growth of human embryonic stem cells[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(3): 237-240. doi: 10.3969/j.issn.1006-0111.2019.03.009
Citation: WANG Zhi, WANG Shuna, XU Tianying, LI Zhiyong, ZHANG Sailong, MIAO Chaoyu. The effect of Nampt gene editing on the growth of human embryonic stem cells[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(3): 237-240. doi: 10.3969/j.issn.1006-0111.2019.03.009
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The effect of Nampt gene editing on the growth of human embryonic stem cells

doi: 10.3969/j.issn.1006-0111.2019.03.009

  • Department of Pharmacology, School of Pharmacy, Naval Medical University, Shanghai 200433, China

  • Received Date: 2018-12-05
  • Rev Recd Date: 2019-04-04
  • Objective To study the role of nicotinamide phosphoribosyltransferase(Nampt)gene in the growth of human embryonic stem cells. Methods CRISPR/Cas9 technology was used for Nampt gene knockout in human embryonic stem cells by introducing frame-shift mutation to gene coding region.Then,the mutational cells were screened with gene sequencing and observed for the growth situation under the microscope.In addition,Nampt inhibitor FK866 was introduced at different concentrations to interfere the growth of human embryonic stem cells,followed by the cell viability test with CCK-8 kit. Results Human embryonic stem cells gradually died after the mutation of Nampt gene with CRISPR/Cas9 technology.Likewise,Nampt inhibitor FK866 significantly inhibited the cell viability and gradually led to the death of cells. Conclusion Nampt gene plays an indispensable role in maintaining the growth of human embryonic stem cells.
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    [2] FATEHULLAH A,TAN S H,BARKER N.Organoids as an in vitro model of human development and disease[J].Nat Cell Biol,2016,18(3):246-254.
    [3] HSU P D,LANDER E S,ZHANG F.Development and applications of CRISPR-cas9 for genome engineering[J].Cell,2014,157(6):1262-1278.
    [4] CONG L,ZHANG F.Genome Engineering Using CRISPR-Cas9 System[J].Methods Mol Biol,2015,1239:197.
    [5] BELENKY P,BOGAN K L,BRENNER C.NAD+ metabolism in health and disease[J].Trends Biochem Sci,2007,32(1):12-19.
    [6] WANG P,MIAO C Y.NAMPT as a therapeutic target against stroke[J].Trends Pharmacol Sci,2015,36(12):891-905.
    [7] XU R,YUAN Z,YANG L,et al.Inhibition of NAMPT decreases cell growth and enhances susceptibility to oxidative stress[J].Oncol Rep,2017,38(3):1767-1773..
    [8] YIN H,VAN DER VEER E,FRONTINI M J,et al.Intrinsic directionality of migrating vascular smooth muscle cells is regulated by NAD+ biosynthesis[J].J Cell Sci,2012,125(23):5770-5780.
    [9] VAN DER VEER E,HO C,O'NEIL C,et al.Extension of human cell lifespan by nicotinamide phosphoribosyltransferase[J].J Biol Chem,2007,282(15):10841-10845.
    [10] STEIN L R,IMAI S I.Specific ablation of Nampt in adult neural stem cells recapitulates their functional defects during aging[J].Embo J,2014,33:1321-1340.
    [11] HENG B C.Effect of Rho-associated kinase (ROCK) inhibitor Y-27632 on the post-thaw viability of cryopreserved human bone marrow-derived mesenchymal stem cells[J].Tissue Cell,2009,41(5):376-380.
    [12] GARTEN A,PETZOLD S,KÖRNER A,et al.Nampt:linking NAD biology,metabolism and cancer[J].Trends Endocrinol Metab,2009,20(3):130-138.
    [13] REVOLLO J R,KÖRNER A,MILLS K F,et al.Nampt/PBEF/Visfatin regulates insulin secretion in β cells as a systemic NAD biosynthetic enzyme[J].Cell Metab,2007,6(5):363-375.
    [14] WANG P,DU H,ZHOU C C,et al.Intracellular NAMPT-NAD+-SIRT1 cascade improves post-ischaemic vascular repair by modulating Notch signalling in endothelial progenitors[J].Cardiovasc Res,2014,104(3):477-488.
    [15] WANG P,XU T Y,GUAN Y F,et al.Nicotinamide phosphoribosyltransferase protects against ischemic stroke through SIRT1-dependent adenosine monophosphate-activated kinase pathway[J].Ann Neurol,2011,69(2):360-374.
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The effect of Nampt gene editing on the growth of human embryonic stem cells

doi: 10.3969/j.issn.1006-0111.2019.03.009
  • Department of Pharmacology, School of Pharmacy, Naval Medical University, Shanghai 200433, China
  • Received Date: 2018-12-05
  • Rev Recd Date: 2019-04-04

Abstract: Objective To study the role of nicotinamide phosphoribosyltransferase(Nampt)gene in the growth of human embryonic stem cells. Methods CRISPR/Cas9 technology was used for Nampt gene knockout in human embryonic stem cells by introducing frame-shift mutation to gene coding region.Then,the mutational cells were screened with gene sequencing and observed for the growth situation under the microscope.In addition,Nampt inhibitor FK866 was introduced at different concentrations to interfere the growth of human embryonic stem cells,followed by the cell viability test with CCK-8 kit. Results Human embryonic stem cells gradually died after the mutation of Nampt gene with CRISPR/Cas9 technology.Likewise,Nampt inhibitor FK866 significantly inhibited the cell viability and gradually led to the death of cells. Conclusion Nampt gene plays an indispensable role in maintaining the growth of human embryonic stem cells.

WANG Zhi, WANG Shuna, XU Tianying, LI Zhiyong, ZHANG Sailong, MIAO Chaoyu. The effect of Nampt gene editing on the growth of human embryonic stem cells[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(3): 237-240. doi: 10.3969/j.issn.1006-0111.2019.03.009
Citation: WANG Zhi, WANG Shuna, XU Tianying, LI Zhiyong, ZHANG Sailong, MIAO Chaoyu. The effect of Nampt gene editing on the growth of human embryonic stem cells[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(3): 237-240. doi: 10.3969/j.issn.1006-0111.2019.03.009
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