PMID- 28155604
OWN - NLM
STAT- MEDLINE
DCOM- 20180529
LR  - 20180924
IS  - 1875-5631 (Electronic)
IS  - 1566-5232 (Linking)
VI  - 16
IP  - 6
DP  - 2017
TI  - A Vector Based on the Chicken Hypersensitive Site 4 Insulator Element Replicates 
      Episomally in Mammalian Cells.
PG  - 410-418
LID - 10.2174/1566523217666170202122755 [doi]
AB  - BACKGROUND: Gene therapy in mammalian cells requires vectors exhibiting long-term 
      stability and high expression. Episomal gene expression vectors offer a safe and 
      attractive alternative to those that integrate into the host cell genome. 
      MATERIALS & METHODS: In the present study, we developed a new episomal vector 
      based on the insulator, chicken hypersensitive site 4 (cHS4). The cHS4 element 
      was artificially synthesized, cloned into the pEGFP-C1 vector, and used to 
      transfect Chinese hamster ovary (CHO) and human Chang liver cells. The stably 
      transfected cell colonies were further cultured in either the presence or absence 
      of G418 selection. Fluorescence in situ hybridization (FISH) analysis and vector 
      rescue experiments demonstrated that the vector replicated episomally in both CHO 
      and human Chang liver cells. Compared with episomal vectors mediated by matrix 
      attachment region sequences, the cHS4 element-containing vector yielded increased 
      transgene expression levels, transfection efficiency, and stability during 
      long-term culture. The vector was present at a very low copy number in the cells 
      and was stably maintained over more than 100 generations without selection 
      pressure. CONCLUSION: In conclusion, apart from a few free vector forms, the 
      cHS4-containing vector mainly replicates episomally in mammalian cells and out- 
      performs comparable systems in terms of yielding both higher expression levels 
      and stability levels.
CI  - Copyright(c) Bentham Science Publishers; For any queries, please email at 
      epub@benthamscience.org.
FAU - Zhang, Xi
AU  - Zhang X
AD  - Department of Biochemistry and Molecular Biology, Xinxiang Medical University, 
      Xinxiang 453003, Henan. China.
FAU - Wang, Xiao-Yin
AU  - Wang XY
AD  - Department of Biochemistry and Molecular Biology, Xinxiang Medical University, 
      Xinxiang 453003, Henan. China.
FAU - Jia, Yan-Long
AU  - Jia YL
AD  - Pharmacy College, Xinxiang Medical University, Xinxiang 453003, Henan. China.
FAU - Guo, Xiao
AU  - Guo X
AD  - Pharmacy College, Xinxiang Medical University, Xinxiang 453003, Henan. China.
FAU - Wang, Yan-Fang
AU  - Wang YF
AD  - Department of Biochemistry and Molecular Biology, Xinxiang Medical University, 
      Xinxiang 453003, Henan. China.
FAU - Wang, Tian-Yun
AU  - Wang TY
AD  - Department of Biochemistry and Molecular Biology, Xinxiang Medical University, 
      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
SB  - IM
MH  - Animals
MH  - CHO Cells
MH  - Chickens/genetics
MH  - Cricetinae
MH  - Cricetulus
MH  - *Genetic Therapy
MH  - Genetic Vectors/genetics/*therapeutic use
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Insulator Elements/*genetics
MH  - Plasmids/genetics/*therapeutic use
MH  - Transfection
OTO - NOTNLM
OT  - Chicken hypersensitive site 4 (cHS4)
OT  - Episomal vectors.
OT  - Insulator element
OT  - Mammalian cells
EDAT- 2017/02/06 06:00
MHDA- 2018/05/31 06:00
CRDT- 2017/02/04 06:00
PHST- 2016/11/30 00:00 [received]
PHST- 2017/01/04 00:00 [revised]
PHST- 2017/01/31 00:00 [accepted]
PHST- 2017/02/06 06:00 [pubmed]
PHST- 2018/05/31 06:00 [medline]
PHST- 2017/02/04 06:00 [entrez]
AID - CGT-EPUB-81449 [pii]
AID - 10.2174/1566523217666170202122755 [doi]
PST - ppublish
SO  - Curr Gene Ther. 2017;16(6):410-418. doi: 10.2174/1566523217666170202122755.