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| The liver X receptor α (LXRα) is a member of the nuclear hormone receptor superfamily which could regulate the transcription of the genes involved in cholesterol transportation. Recent studies showed that LXRα is considered as a critical regulator in cholesterol homeostasis in macrophages, which could regulate several genes involved in cholesterol transport, such as the ATP-binding cassette trans-porters (ABCs), ABCA1, ABCG1, apolipoprotein E (ApoE) and lipoprotein lipase (LPL). In our study, four pairs of inhibition shRNA were designed to exclusively target bovine LXRα mRNA. Lentiviral vector containing LXRα shRNAs were constructed through BP and LP recombination system, and used to obtained the corresponding lentiviruses in 293T cells. The titers of the Lenti-14MR0054-01, -2, -3 and -04 viruses were 3×108TU/ml, 2×108TU/ml, 2.5×108TU/ml, 3.5×108TU/ml, respectively. The knockdown efficiency of pLenti-03 targeting LXRα reached 88 % in bovine muscle satellite cells. Furthermore, the mRNA expression of RXRα was upregulated in bovine muscle satellite cells, whereas that of PPARα, PPARγ, ABCA1, LPL, ApoE were downregulated at 48 h post-pLenti-03 viruses infection. Therefore, the efficient lentivirus vector inhibiting bovine LXRα expression was obtained in the present study, providing basis for loss-of-function of LXRα in bovine muscle satellite cells |
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| Keywords:bovine LXRα gene; lentivirus; vector construction; gene silencing |
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