Preparation of Recombinant Plasmid DNA for DNA-Mediated Gene Transfer

Author(s):  
Rémy Aubin ◽  
Michael Weinfeld ◽  
Malcolm C. Paterson
1995 ◽  
Vol 268 (6) ◽  
pp. L1052-L1056 ◽  
Author(s):  
M. F. Tsan ◽  
J. E. White ◽  
B. Shepard

A number of gene delivery methods have been developed to facilitate gene transfer into mammalian somatic cells in vivo. In this study, we demonstrated that tracheal insufflation of two recombinant plasmids containing a bacterial gene, chloramphenicol acetyltransferase (CAT), driven by the cytomegalovirus (CMV) immediate early promoter/enhancer, either alone or complexed to cationic liposomes, resulted in efficient and selective transfection of the lungs in rats. When the simian virus 40 (SV40) promoter/enhancer was used, there was no detectable transfection. Insufflation of plasmid DNA was as efficient as plasmid-liposome complexes in transfecting the lungs. Expression of the CAT gene in the lungs was noted as early as 1 day after insufflation of plasmid DNA alone or plasmid-liposome complexes, and lasted for > 21 days. In contrast, intravenous injection of plasmid alone or plasmid-liposome complexes did not result in transfection of the lungs. Because of its simplicity, without the potential adverse effect of any gene delivery systems, intratracheal delivery of recombinant plasmid DNA may have potential implication for lung-specific gene therapy.


2006 ◽  
Vol 32 (5) ◽  
pp. 449-453 ◽  
Author(s):  
Pierre Collinet ◽  
Rodolphe Vereecque ◽  
Frédéric Sabban ◽  
Denis Vinatier ◽  
Eric Leblanc ◽  
...  

1991 ◽  
Vol 71 (2) ◽  
pp. 287-291 ◽  
Author(s):  
J. S. Gavora ◽  
B. Benkel ◽  
H. Sasada ◽  
W. J. Cantwell ◽  
P. Fiser ◽  
...  

Experiments were carried out to transform laboratory mice and domestic chickens by use of sperm incubated with bacterial plasmid DNA. Following demonstration of "uptake" of such DNA by sperm of both species, attempts were made to replicate a previously published procedure (Lavitrano et al. 1989, Cell 57: 717–723) for producing transgenic mice through in vitro fertilization (IVF). Also, female mice and hens were inseminated (AI) with sperm which had been incubated in a DNA solution. Such incubation did not influence the fertility or hatchability of the hens' eggs. However, no transformed progeny were detected among 45 mice produced by IVF or among 69 mice and 470 chickens produced by AI. Key words: Sperm-mediated DNA transfer, mice, chickens


BioTechniques ◽  
1997 ◽  
Vol 23 (4) ◽  
pp. 603-608 ◽  
Author(s):  
Biao Li ◽  
Kirsten Y. Pilcher ◽  
Theodore E. Wyman ◽  
Curtis A. Machida

2010 ◽  
Vol 2010 ◽  
pp. 1-10 ◽  
Author(s):  
Syahril Abdullah ◽  
Wai Yeng Wendy-Yeo ◽  
Hossein Hosseinkhani ◽  
Mohsen Hosseinkhani ◽  
Ehab Masrawa ◽  
...  

A novel cationic polymer, dextran-spermine (D-SPM), has been found to mediate gene expression in a wide variety of cell lines andin vivothrough systemic delivery. Here, we extended the observations by determining the optimal conditions for gene expression of D-SPM/plasmid DNA (D-SPM/pDNA) in cell lines and in the lungs of BALB/c mice via instillation delivery.In vitrostudies showed that D-SPM could partially protect pDNA from degradation by nuclease and exhibited optimal gene transfer efficiency at D-SPM to pDNA weight-mixing ratio of 12. In the lungs of mice, the levels of gene expression generated by D-SPM/pDNA are highly dependent on the weight-mixing ratio of D-SPM to pDNA, amount of pDNA in the complex, and the assay time postdelivery. Readministration of the complex at day 1 following the first dosing showed no significant effect on the retention and duration of gene expression. The study also showed that there was a clear trend of increasing size of the complexes as the amount of pDNA was increased, where the sizes of the D-SPM/pDNA complexes were within the nanometer range.


2004 ◽  
Vol 1274 ◽  
pp. 159-163
Author(s):  
Yoshiaki Taniyama ◽  
Munehisa Shimamura ◽  
Naoyuki Sato ◽  
Naruya Tomita ◽  
Masayuki Endho ◽  
...  

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