Generation of mice with hepatocyte-specific conditional deletion of sphingosine kinase 1.

Transgenic Research
Jinfeng YuTian Lan

Abstract

SphK1 gene has different roles in various types of cells in liver diseases, but most studies are based on global knockout mice, which hampers the study on the cellular and molecular mechanisms of SphK1. In order to further study the role of SphK1 in liver, SphK1 conditional knockout mice were constructed. A liver-specific SphK1 gene knockout mouse model was constructed by the Cre/Loxp recombinant enzyme system. PCR technologies and western blotting were used to identified the elimination of SphK1 gene in hepatocytes. SphK1flox/flox mice were used as a control group to verify the effectiveness of SphK1 liver-specific knockout mice from the profile, pathology, and serology of mice. The ablation of SphK1 in hepatic parenchymal cells was demonstrated by fluorescent in situ hybridization and the contents of S1P and Sph were measured by ELISA kit. The genotypes of liver in SphK1 conditional knockout mice were different from that of other organs. The mRNA and protein levels of SphK1 in liver tissue of SphK1 conditional knockout mice were almost depleted by compared with SphK1flox/flox mice. Physiology and pathology showed no significant difference between SphK1 liver conditional knockout mice and SphK1flox/flox mice. Additionally, Sph...Continue Reading

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Citations

Feb 27, 2021·Frontiers in Endocrinology·Yanfei QiPu Xia

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Methods Mentioned

BETA
genetic modification
gene knockout
transgenic
PCR
genotyping
reverse transcription PCR
electrophoresis
ELISA

Software Mentioned

GraphPad Prism

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