PMID- 25295466
OWN - NLM
STAT- MEDLINE
DCOM- 20160204
LR  - 20150209
IS  - 1533-4023 (Electronic)
IS  - 0160-2446 (Linking)
VI  - 65
IP  - 2
DP  - 2015 Feb
TI  - Gene therapy for hemophilia B with liver-specific element mediated by Rep-RBE 
      site-specific integration system.
PG  - 153-9
LID - 10.1097/FJC.0000000000000172 [doi]
AB  - Adeno-associated virus (AAV) is a nonpathogenic virus capable of targeting human 
      chromosome 19 for integration at AAVS1 site, and a 16 bp Rep binding element 
      (RBE) sequence of AAV was sufficient for mediating this specific integration in 
      the presence of AAV regulation proteins (Rep). Previously, we cotransduced 2 
      plasmids, pRBE-CMV-hFIX and pRC, into the AAVS1 transgenic mice by hydrodynamic 
      injection, and a long-term expression of human coagulation Factor IX (hFIX) was 
      observed. The corresponding AAVS1 locus site-specific integrations were verified 
      by nested polymerase chain reaction. In this study, we established a novel hFIX 
      expression plasmid, pRBE-HCR-hAAT-hFIX, driven by a liver-specific promoter by 
      replacing the CMV promoter of pRBE-CMV-hFIX with a humanized promoter consisting 
      of HCR-hAAT. The expression of hFIX in vitro was almost the same in transient 
      transfection of pRBE-CMV-hFIX or pRBE-HCR-hAAT-hFIX. AAVS1-specific integrations 
      were identified both in mice transfected with pRC/pRBE-CMV-hFIX cocktail and 
      pRC/pRBE-HCR-hAAT-hFIX cocktail. However, the expression of hFIX of 
      pRBE-HCR-hAAT-hFIX mice was higher and persisted longer. It achieved more than 1% 
      of normal plasma hFIX concentration and maintained for 240 days. The result 
      suggested that RBE-HCR-hAAT element could improve the expression of hFIX and 
      present potential usage of Rep-RBE site-specific integration in gene therapy for 
      hemophilia B.
FAU - Xu, Zhengxin
AU  - Xu Z
AD  - *State Key Laboratory of Genetic Engineering, Institute of Genetics, School of 
      Life Sciences, Fudan University, Shanghai, China; and daggerDepartment of 
      Pharmacology, Medical School, Yangzhou University, Yangzhou, China.
FAU - Ye, Juan
AU  - Ye J
FAU - Zhang, Amin
AU  - Zhang A
FAU - Xie, Linjun
AU  - Xie L
FAU - Shen, Qi
AU  - Shen Q
FAU - Xue, Jinglun
AU  - Xue J
FAU - Chen, Jinzhong
AU  - Chen J
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Cardiovasc Pharmacol
JT  - Journal of cardiovascular pharmacology
JID - 7902492
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Viral Proteins)
RN  - 137750-19-7 (rep proteins, Adeno-associated virus 2)
RN  - 9001-28-9 (Factor IX)
SB  - IM
MH  - Animals
MH  - Cells, Cultured
MH  - *Chromosomes, Human, Pair 19
MH  - DNA-Binding Proteins/*genetics
MH  - Dependovirus/physiology
MH  - Factor IX/genetics
MH  - Gene Transfer Techniques
MH  - Genetic Therapy/*methods
MH  - Genetic Vectors
MH  - Hemophilia B/*therapy
MH  - Humans
MH  - Liver/metabolism
MH  - Mice
MH  - Mice, Transgenic
MH  - Plasmids/physiology
MH  - Promoter Regions, Genetic
MH  - Viral Proteins/*genetics
EDAT- 2014/10/09 06:00
MHDA- 2016/02/05 06:00
CRDT- 2014/10/09 06:00
PHST- 2014/10/09 06:00 [entrez]
PHST- 2014/10/09 06:00 [pubmed]
PHST- 2016/02/05 06:00 [medline]
AID - 10.1097/FJC.0000000000000172 [doi]
PST - ppublish
SO  - J Cardiovasc Pharmacol. 2015 Feb;65(2):153-9. doi: 10.1097/FJC.0000000000000172.