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Expression of an Episomal Vector in Developing Chicken Embryo and Chicks
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作者 Mu Yan-shuang Wang Jia-qiang +3 位作者 Yin Zhi Jiang Dan-dan Li Hui Liu Zhong-hua 《Journal of Northeast Agricultural University(English Edition)》 CAS 2013年第3期17-24,F0003,共9页
The domesticated chicken has important roles in basic and applied research. The vector based on scaffold matrix attachment region (S/MAR) appears to be sufficient to maintain long-term expression in an episomal stat... The domesticated chicken has important roles in basic and applied research. The vector based on scaffold matrix attachment region (S/MAR) appears to be sufficient to maintain long-term expression in an episomal state in various mammalian cells. To explore the practical use of episomal vector in transgene technology of agricultural chickens, we fused the S/MAR of the human r-interferon gene into the pEGFP vector and transduced it into chickens by sperm-mediated gene transfer (SMGT). PCR detection indicated the positive rate of transgene chickens was 60%. The RT-PCR detection and fluorescence observation confirmed the expression of the GFP and indicated the existence of the GFP during the chicken embryogenesis and fetal development. The PCR detection and rescue experiments confirmed the episomal state of the pEPI-EGFP in chick embryos and chicks. These results showed that the S/MAR-based vector could function properly in chicken embryos and was a practicable tool combined with the SMGT to study the development of chickens. 展开更多
关键词 TRANSGENE scaffold/matrix attachment region sperm-mediated gene transfer CHICKEN
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Non-integrating lentiviral vectors based on the minimal S/MAR sequence retain transgene expression in dividing cells 被引量:1
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作者 Zhen Xu Feng Chen +7 位作者 Lingling Zhang Jing Lu Peng Xu Guang Liu Xuemin Xie Wenli Mu Yajun Wang Depei Liu 《Science China(Life Sciences)》 SCIE CAS CSCD 2016年第10期1024-1033,共10页
Safe and efficient gene transfer systems are the basis of gene therapy applications. Non-integrating lentiviral(NIL) vectors are among the most promising candidates for gene transfer tools, because they exhibit high t... Safe and efficient gene transfer systems are the basis of gene therapy applications. Non-integrating lentiviral(NIL) vectors are among the most promising candidates for gene transfer tools, because they exhibit high transfer efficiency in both dividing and non-dividing cells and do not present a risk of insertional mutagenesis. However, non-integrating lentiviral vectors cannot introduce stable exogenous gene expression to dividing cells, thereby limiting their application. Here, we report the design of a non-integrating lentiviral vector that contains the minimal scaffold/matrix attachment region(S/MAR) sequence(SNIL), and this SNIL vector is able to retain episomal transgene expression in dividing cells. Using SNIL vectors, we detected the expression of the eGFP gene for 61 days in SNIL-transduced stable CHO cells, either with selection or not. In the NIL group without the S/MAR sequence, however, the transduced cells died under selection for the transient expression of NIL vectors. Furthermore,Southern blot assays demonstrated that the SNIL vectors were retained extrachromosomally in the CHO cells. In conclusion,the minimal S/MAR sequence retained the non-integrating lentiviral vectors in dividing cells, which indicates that SNIL vectors have the potential for use as a gene transfer tool. 展开更多
关键词 gene transfer non-integrating lentivirus scaffold/matrix attachment region (S/MAR) episomal vectors
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