Intranuclear disposition of exogenous DNA in vivo: silencing, methylation and fragmentation.
The intranuclear disposition of exogenous DNA is highly important for the therapeutic effects of the administrated DNA. Naked luciferase-plasmid DNA was delivered into mouse liver by a hydrodynamics-based injection, and the amounts of intranuclear plasmid DNA, luciferase, and its mRNA were quantitated at various time points. Methylation of the promoter of the luciferase gene was also analyzed. Expression efficiency from one copy of the exogenous DNA dramatically decreased over time, and the DNA was methylated and degraded into fragments. Unexpectedly, methylation of the intact plasmid DNA was low and did not increase over time. Rather, the fragmented DNA was methylated more frequently than the intact plasmid. These results suggest that the CpG methylation and the degradation of exogenous DNA, and its 'silencing', occurred in parallel in the nucleus.
Ochiai
Hiroshi
H
Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Sapporo 060-0812, Japan.
Harashima
Hideyoshi
H
Kamiya
Hiroyuki
H
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D018899:CpG Islands / Q000502:physiology; D053938:DNA Fragmentation / Q000502:physiology*; D019175:DNA Methylation; D005260:Female; D020868:Gene Silencing / Q000502:physiology*; D007275:Injections, Intravenous; D008099:Liver / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000008:administration & dosage / Q000378:metabolism* / Q000627:therapeutic use; D011401:Promoter Regions, Genetic / Q000235:genetics
D008156:Luciferases