PMID- 27915978
OWN - NLM
STAT- MEDLINE
DCOM- 20180116
LR  - 20180818
IS  - 1875-5631 (Electronic)
IS  - 1566-5232 (Linking)
VI  - 16
IP  - 4
DP  - 2016
TI  - Cell Compatibility of an Eposimal Vector Mediated by the Characteristic Motifs of 
      Matrix Attachment Regions.
PG  - 271-277
LID - 10.2174/1566523216666161202160936 [doi]
AB  - The characteristic sequence of beta-interferon matrix attachment regions (MARs) can 
      mediate transgene expression via episomal vectors in Chinese hamster ovary (CHO) 
      cells. However, the host cells were from hamster ovaries, which are not suitable 
      target cells for gene therapy. In this study, we aimed to evaluate the 
      suitability of 12 different human cell lines as target cells for gene therapy. We 
      transfected the cells with episomal vectors and obtained colonies stably 
      expressing the vector products after G418 screening. Therefore the stably 
      transfected cells were split into two and further cultured either in the presence 
      or the absence of G418. Flow cytometry was used to observe the positive rate of 
      cell transfection and level of green fluorescent protein (GFP) expression. 
      Plasmid rescue assays, fluorescence in situ hybridization (FISH), and 
      fluorescence quantitative polymerase chain reaction (PCR) were used to 
      investigate the presence and gene copy numbers of plasmid in mammalian cells. The 
      results showed that transfection efficiency and transgene expression levels in 
      A375, Eca-109, and Changliver cells were high. In contrast, transgene silencing 
      was observed in BJ, HSF, and A431 cells, and low expression of the transgene was 
      observed in the other six cell lines. In addition, the plasmid was present in the 
      episomal state in A375, Eca-109, and Chang-liver cells with relatively low copy 
      numbers even under nonselective conditions. Thus, our results provide the first 
      evidence showing transgene expression of an episomal vector mediated by the 
      characteristic motifs of MARs for maintenance of the longterm stability of 
      episomes in different types of cells.
CI  - Copyright(c) Bentham Science Publishers; For any queries, please email at 
      epub@benthamscience.org.
FAU - Wang, Tian-Yun
AU  - Wang TY
FAU - Wang, Li
AU  - Wang L
FAU - Yang, Yu-Xin
AU  - Yang YX
FAU - Zhao, Chun-Peng
AU  - Zhao CP
FAU - Jia, Yan-Long
AU  - Jia YL
FAU - Li, Qin
AU  - Li Q
FAU - Zhang, Jun-He
AU  - Zhang JH
FAU - Peng, Yi-You
AU  - Peng YY
FAU - Wang, Miao
AU  - Wang M
FAU - Xu, Hong-Yan
AU  - Xu HY
FAU - Wang, Xiao-Yin
AU  - Wang XY
AD  - Department of Biochemistry and Molecular Biology, Xinxiang Medical University, 
      Jinsui Road, Xinxiang 453003, Henan, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United Arab Emirates
TA  - Curr Gene Ther
JT  - Current gene therapy
JID - 101125446
RN  - 0 (Recombinant Proteins)
RN  - 0 (enhanced green fluorescent protein)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Gene Dosage
MH  - Genetic Vectors/*genetics
MH  - Green Fluorescent Proteins/genetics/metabolism
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Matrix Attachment Regions/*genetics
MH  - Plasmids/genetics
MH  - Recombinant Proteins/genetics/*metabolism
MH  - Transfection/methods
OTO - NOTNLM
OT  - Eposimal vector
OT  - Host cell
OT  - Matrix attachment regions
OT  - Transgene expression
EDAT- 2016/12/06 06:00
MHDA- 2018/01/18 06:00
CRDT- 2016/12/06 06:00
PHST- 2016/05/20 00:00 [received]
PHST- 2016/11/02 00:00 [revised]
PHST- 2016/11/18 00:00 [accepted]
PHST- 2016/12/06 06:00 [pubmed]
PHST- 2018/01/18 06:00 [medline]
PHST- 2016/12/06 06:00 [entrez]
AID - CGT-EPUB-80118 [pii]
AID - 10.2174/1566523216666161202160936 [doi]
PST - ppublish
SO  - Curr Gene Ther. 2016;16(4):271-277. doi: 10.2174/1566523216666161202160936.