Silenciamiento de la fosfoglucosa isomerasa (PGI) en fibroblastos inmortalizados de ratón. Mediante la técnica del RNA de interferencia

  1. FERNANDEZ-SOBERON, Silvia 1
  2. BOLAÑOS, Juan Pedro 1
  1. 1 Institute of Functional Biology and Genomics (IBFG), University of Salamanca-CSIC, Spain.
Aldizkaria:
Farmajournal

ISSN: 2445-1355

Argitalpen urtea: 2016

Alea: 1

Zenbakia: 2

Orrialdeak: 57-65

Mota: Artikulua

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Beste argitalpen batzuk: Farmajournal

Laburpena

A target-specific small hairpin RNA (shRNA) was added into a plasmid delivery system (pSUPER-NeoGFP) and introduced into a cell culture of mouse embryonic fibroblasts (MEFs). The shRNA was constructed for suppressing   the expression in mice cells glucose-6-phosphate isomerase PGIm. Glucose-6-phosphate isomerase is an enzyme that catalyses the conversion of glucose-6-phosphate into fructose 6-phosphate in glycolysis’ second step. A PEI–polyethylene glycol conjugate was used as a pSUPER-NeoGFP-shPGI plasmid carrier. Two cultures of cells were transformed one used as control with the plasmid delivery system (pSUPER-NeoGFP) and the other with the plasmid with the sh-PGI. The gene inhibition and cellular uptake behaviours were explored by the measurement of lactic acid as a marker of the efficiency of glycolysis and with a reverse transcription polymerase chain reaction (RT-PCR) to quantify PGIm expresion. No significatively differences were found in the lactic acid production while a decrease in the PGIm expresion was observed.

Erreferentzia bibliografikoak

  • Nelson, David L.; Michael M. Cox: Lehninger principles of biochemistry. Vol 1.5th ed. New York: W. H. Freeman and Company, cop. 2008.
  • Reynolds, Angela, et al. Rational siRNA design for RNA interference. NATURE BIOTECHNOL, 2004, vol. 22, no 3, p. 326-330.
  • Rodriguez-Rodrigez, Patricia; Fernández, Emilio; Bolaños, Juan P. Underestimation of the pentose–phosphate pathway in intact primary neurons as revealed by metabolic flux analysis. J CEREBR BLOOD F MET, 2013, vol. 33, no 12, p. 1843-1845.
  • Voet D, Voet Jc, Pratt, CW. Fundamentals of biochemistry. 5th ed. New York: John Wiley and Sons; 1998.
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