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Information Journal Paper

Title

EFFECTS OF KLF4 AND C-MYC KNOCKDOWN ON PLURIPOTENCY MAINTENANCE IN PORCINE INDUCED PLURIPOTENT STEM CELL

Pages

  640-646

Abstract

 Objective: The importance of Oct4 and Sox2 in maintaining PLURIPOTENCY and self-renewal is well-understood, but the functions ofKlf4 and C-MYC has not been fully investigated. In the present study, we attempted to determine the roles of KLF4 and C-MYC on PLURIPOTENCY maintenance of porcine induced pluripotent stem (piPS) cells.Materials and Methods: In this experimental study, we performed SHORT HAIRPIN RNA (shRNA) to knock down the KLF4and C-MYC functions of piPS cells and examined PLURIPOTENCY markers and teratoma formation to evaluate piPS cell PLURIPOTENCY. The shRNA-Klf4 and shRNA-C-MYC vectors containing a reporter gene, TagFP635, were transfected into piPS cells by lentivirus infection. The piPS cells fully expressing infrared fluorescence were selected to confirm gene knockdown ofKlf4 and C-MYC reverse transcription-polymerase chain reaction (RT-PCR). Next, for PLURIPOTENCY evaluation, expression of PLURIPOTENCY markers was detected by immunocytochemical staining, and capability of teratoma formation was investigated by piPS cell transplantation into nonobese diabetic-severe combined immunodeficiency (NOD-SCID) mice.Results: Our findings indicated that KLF4 and C-MYC functions of piPS cells were knocked down by shRNA transfection, and knockdown ofKlf4 and C-MYC functions impaired expression of PLURIPOTENCY markers such as Oct4, AP, SSEA-3, SSEA-4, TRA-1-6, and TRA-1-81. Furthermore, piPS cells withoutKlf4 and C-MYC expression failed to form teratomas.Conclusion: The PLURIPOTENCY of piPS cells are crucially dependent upon KLF4 and C-MYC expression. These findings, suggesting potential mechanisms ofKlf4 and C-MYC contribution to piPS cell formation, have important implications for application, regulation, and tumorigenesis of piPS cells.

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    APA: Copy

    LIAO, YU JING, CHEN, YI SHIOU, LEE, JA XIN, CHEN, LIH REN, & YANG, JENN RONG. (2018). EFFECTS OF KLF4 AND C-MYC KNOCKDOWN ON PLURIPOTENCY MAINTENANCE IN PORCINE INDUCED PLURIPOTENT STEM CELL. CELL JOURNAL (YAKHTEH), 19(4), 640-646. SID. https://sid.ir/paper/699067/en

    Vancouver: Copy

    LIAO YU JING, CHEN YI SHIOU, LEE JA XIN, CHEN LIH REN, YANG JENN RONG. EFFECTS OF KLF4 AND C-MYC KNOCKDOWN ON PLURIPOTENCY MAINTENANCE IN PORCINE INDUCED PLURIPOTENT STEM CELL. CELL JOURNAL (YAKHTEH)[Internet]. 2018;19(4):640-646. Available from: https://sid.ir/paper/699067/en

    IEEE: Copy

    YU JING LIAO, YI SHIOU CHEN, JA XIN LEE, LIH REN CHEN, and JENN RONG YANG, “EFFECTS OF KLF4 AND C-MYC KNOCKDOWN ON PLURIPOTENCY MAINTENANCE IN PORCINE INDUCED PLURIPOTENT STEM CELL,” CELL JOURNAL (YAKHTEH), vol. 19, no. 4, pp. 640–646, 2018, [Online]. Available: https://sid.ir/paper/699067/en

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