PMID- 12795598
OWN - NLM
STAT- MEDLINE
DCOM- 20030724
LR  - 20061115
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 42
IP  - 23
DP  - 2003 Jun 17
TI  - Functional characterization of transgene integration patterns by halo 
      fluorescence in situ hybridization: electroporation versus retroviral infection.
PG  - 7035-43
AB  - Gene transfer techniques are essential for bioengineering and gene therapy. 
      Retroviral vectors constitute an important tool in this context as 
      integrase-catalyzed genomic anchoring is one of the best defined results of 
      nonhomologous recombination, occurring at loci with favorable expression 
      properties. On the basis of a retroviral expression cassette, this study focuses 
      on differences regarding the genomic targets after its transfer by either 
      retroviral infection or electroporation. Fluorescence in situ hybridization 
      (FISH) of clones generated by infection revealed for the majority an association 
      of the transgene with the nuclear matrix while this was true only for a minority 
      of single copy clones if the same construct was transferred by electroporation. 
      Our results demonstrate that the process of integration is distinct for both gene 
      transfer routes and confirm that retroviral survival strategies favor integration 
      next to scaffold/matrix attachment regions. These results may be one key to 
      explain recent consequences of gene therapy trials that have led to the 
      deregulation of endogenous genes.
FAU - Goetze, S
AU  - Goetze S
AD  - GBF-German Research Center for Biotechnology/Epigenetic Regulation, Mascheroder 
      Weg 1, D-38124 Braunschweig, Germany.
FAU - Huesemann, Y
AU  - Huesemann Y
FAU - Baer, A
AU  - Baer A
FAU - Bode, J
AU  - Bode J
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Cell Nucleus/chemistry
MH  - Cricetinae
MH  - Electroporation
MH  - *Gene Transfer Techniques
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Nuclear Matrix/genetics/metabolism
MH  - Plasmids/metabolism
MH  - Regulatory Sequences, Nucleic Acid
MH  - Retroviridae/*genetics
MH  - Transcription, Genetic/genetics
MH  - Transfection
MH  - *Transgenes
MH  - Virus Integration/*genetics
EDAT- 2003/06/11 05:00
MHDA- 2003/07/25 05:00
CRDT- 2003/06/11 05:00
PHST- 2003/06/11 05:00 [pubmed]
PHST- 2003/07/25 05:00 [medline]
PHST- 2003/06/11 05:00 [entrez]
AID - 10.1021/bi0340907 [doi]
PST - ppublish
SO  - Biochemistry. 2003 Jun 17;42(23):7035-43. doi: 10.1021/bi0340907.