Non-integrating lentiviral vectors based on the minimal S/MAR sequence retain transgene expression in dividing cells  

作　　者：Zhen Xu Feng Chen Lingling Zhang Jing Lu Peng Xu Guang Liu Xuemin Xie Wenli Mu Yajun Wang Depei Liu 

机构地区：[1]State Key Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences and Peking Union Medical College

出　　处：《Science China(Life Sciences)》2016年第10期1024-1033,共10页中国科学（生命科学英文版）

基　　金：supported by the Major State Basic Research Development Program of China(2011CB965203)

摘　　要：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.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 in- troduce 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 expres- sion 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. Further- more, Southern blot assays demonstrated that the SNIL vectors were retained extrachromosomally in the CHO cells. In con- clusion, 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.

关 键 词：gene transfer non-integrating lentivirus scaffold/matrix attachment region （S/MAR） episomal vectors 

分 类 号：R450[医药卫生—治疗学]