Establishment of fumarylacetoacetate hydrolase gene knockout rat embryonic stem cell strains

doi:10.3969/j.issn.2095-4344.2013.01.008

Abstract

Abstract:

BACKGROUND: Mouse liver injury model has been widely used. The pathological mechanism of liver injury and the environment of liver transplantation in rats can be used for simulation in humans. Establishing fuarylacetoacetat hydrolase gene knockout embryonic stem cell strains is the key to establish a rat model of liver injury.
OBJECTIVE: To establish fuarylacetoacetat hydrolase gene knockout rat embryonic stem cell strains.
METHODS: We first constructed the targeting vector in vitro with positive-negative selective cassette, then transfered the linearized vector into embryonic stem cells through electroporation. We used embryonic stem cell culture medium containing 0.5 μg/mL puromycin to select puromycin resistant clones 36 hours after electroporation, and then we performed PCR to confirm targeted embryonic stem cell clones.
RESULTS AND CONCLUSION: After electroporation, about 90% of embryonic stem cell clones expressed green fluorescence. Two embryonic stem cell clones expressing green fluorescence were selected and confirmed by PCR. The fuarylacetoacetat hydrolase gene deficient rat embryonic stem cell strains were successfully established. This provides evidence for later establishing gene knockout animal models.

Key words: stem cells, embryonic stem cells, gene knockout, homologous recombination, Fah gene, rats, stem cell photographs-containing paper

CLC Number:

R394.2

Ye Zhen-long, Wang Yan, Jin Hua-jun, Liu Tao, Qian Qi-jun. Establishment of fumarylacetoacetate hydrolase gene knockout rat embryonic stem cell strains[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(1): 50-55.

Figures/Tables 4

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