PMID- 16595609
OWN - NLM
STAT- MEDLINE
DCOM- 20061003
LR  - 20121115
IS  - 0964-6906 (Print)
IS  - 0964-6906 (Linking)
VI  - 15
IP  - 10
DP  - 2006 May 15
TI  - A highly functional mini-dystrophin/GFP fusion gene for cell and gene therapy 
      studies of Duchenne muscular dystrophy.
PG  - 1610-22
AB  - A promising approach for treating Duchenne muscular dystrophy (DMD) is by 
      autologous cell transplantation of myogenic stem cells transduced with a 
      therapeutic expression cassette. Development of this method has been hampered by 
      a low frequency of cellular engraftment, the difficulty of tracing transplanted 
      cells, the rapid loss of autologous cells carrying marker genes that are unable 
      to halt muscle necrosis and the difficulty of stable transfer of a large 
      dystrophin gene into myogenic stem cells. We engineered a 5.7 kb miniDys-GFP 
      fusion gene by replacing the dystrophin C-terminal domain (DeltaCT) with an eGFP 
      coding sequence and removing much of the dystrophin central rod domain 
      (DeltaH2-R19). In a transgenic mdx(4Cv) mouse expressing the miniDys-GFP fusion 
      protein under the control of a skeletal muscle-specific promoter, the green 
      fusion protein localized on the sarcolemma, where it assembled the 
      dystrophin-glycoprotein complex and completely prevented the development of 
      dystrophy in transgenic mdx(4Cv) muscles. When myogenic and other stem cells from 
      these mice were transplanted into mdx(4Cv) recipients, donor cells can be readily 
      identified in skeletal muscle by direct green fluorescence or by using antibodies 
      against GFP or dystrophin. In mdx(4Cv) mice reconstituted with bone marrow cells 
      from the transgenic mice, we monitored engraftment in various muscle groups and 
      found the number of miniDys-GFP(+) fibers increased with time. We suggest that 
      these transgenic mdx(4Cv) mice are highly useful for developing autologous cell 
      therapies for DMD.
FAU - Li, Sheng
AU  - Li S
AD  - Department of Neurology, University of Washington School of Medicine, Seattle, 
      98195-7720, USA.
FAU - Kimura, En
AU  - Kimura E
FAU - Ng, Rainer
AU  - Ng R
FAU - Fall, Brent M
AU  - Fall BM
FAU - Meuse, Leonard
AU  - Meuse L
FAU - Reyes, Morayma
AU  - Reyes M
FAU - Faulkner, John A
AU  - Faulkner JA
FAU - Chamberlain, Jeffrey S
AU  - Chamberlain JS
LA  - eng
GR  - P01 AG015434/AG/NIA NIH HHS/United States
GR  - P01 AS046788/AS/ASC OASH HHS/United States
GR  - P01 NS46788/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20060404
PL  - England
TA  - Hum Mol Genet
JT  - Human molecular genetics
JID - 9208958
RN  - 0 (Dystrophin)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (enhanced green fluorescent protein)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Dystrophin/biosynthesis/*genetics
MH  - *Genetic Therapy
MH  - Green Fluorescent Proteins/*genetics
MH  - Mice
MH  - Mice, Transgenic
MH  - Muscle Contraction
MH  - Muscle Fibers, Skeletal/metabolism/pathology
MH  - Muscle, Skeletal/metabolism/pathology/physiopathology
MH  - Muscular Dystrophy, Duchenne/*genetics/metabolism/therapy
MH  - Promoter Regions, Genetic
MH  - Protein Structure, Tertiary
MH  - Recombinant Fusion Proteins/biosynthesis/*genetics
MH  - Sarcolemma/metabolism
MH  - Stem Cell Transplantation
EDAT- 2006/04/06 09:00
MHDA- 2006/10/04 09:00
CRDT- 2006/04/06 09:00
PHST- 2006/04/06 09:00 [pubmed]
PHST- 2006/10/04 09:00 [medline]
PHST- 2006/04/06 09:00 [entrez]
AID - ddl082 [pii]
AID - 10.1093/hmg/ddl082 [doi]
PST - ppublish
SO  - Hum Mol Genet. 2006 May 15;15(10):1610-22. doi: 10.1093/hmg/ddl082. Epub 2006 Apr 
      4.