Construction of Nanog lentiviral expression vector and its application in differentiation regulation of embryonic stem cells
Chen Fei
Wan Li
Li Yan
Li Xin
Abstract：BACKGROUND:There is a lack of safe and effective modular lentivectors in differentiation regulation of embryonic stem cel s. OBJECTIVE:To construct a modular lentivector, Puro-Nanog-hrGFP, with Nanog promoter-control ed humanized renil a reniformis green fluorescent protein (hrGFP) and puromycin and to observe the expression of Nanog during the differentiation of Nanog-hrGFP-transfected mouse embryonic stem cel lines. METHODS:After PCR amplification, Nanog, hrGFP and entry vector were recombined into the pDest-puro vectors to generate the lentiviral expression vector, Puro-Nanog-hrGFP, by the LR reaction. Through lentivirus production, transduction and puromycin screening, the transduced cel lines with Nanog-hrGFP gene were generated and identified. Expression of Nanog during the differentiation of transgenic mouse cel lines was detected. RESULTS AND CONCLUSION:The lentiviral expression vectors Puro-Nanog-hrGFP were constructed successful y by Gateway technology, and then the transducted cel lines were obtained by lentiviral infection. The expression of Nanog was gradual y decreased during the process of transgenic cel lines differentiation, which provides a new tool for further investigation on regulation of stem cel differentiation.
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Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 2763-2769 )