Intranuclear disposition of exogenous DNA in vivo: Silencing, methylation and fragmentation

被引：39

作者：

Ochiai, H ^{[1
]}

Harashima, H ^{[1
]}

Kamiya, H ^{[1
]}

机构：

[1] Hokkaido Univ, Grad Sch Pharmaceut Sci, Kita Ku, Sapporo, Hokkaido 0600812, Japan

来源：

FEBS LETTERS | 2006年 / 580卷 / 03期

基金：

日本学术振兴会;

关键词：

exogenous DNA; intranuclear disposition; silencing; methylation; fragmentation; hydrodynamics-based injection;

D O I：

10.1016/j.febslet.2006.01.017

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (c) 2006 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

引用

页码：918 / 922

页数：5

共 26 条

[1] METHYLATION AND IMPRINTING - FROM HOST DEFENSE TO GENE-REGULATION [J].

BARLOW, DP .

SCIENCE, 1993, 260 (5106) :309-310

[2] DNA METHYLATION INHIBITS TRANSCRIPTION INDIRECTLY VIA A METHYL-CPG BINDING-PROTEIN [J].

BOYES, J ;

BIRD, A .

CELL, 1991, 64 (06) :1123-1134

[3] Transcriptional silencing is associated with extensive methylation of the CMV promoter following adenoviral gene delivery to muscle [J].

Brooks, AR ;

Harkins, RN ;

Wang, PY ;

Qian, HS ;

Liu, PX ;

Rubanyi, GM .

JOURNAL OF GENE MEDICINE, 2004, 6 (04) :395-404

[4] DNA METHYLATION AND GENE ACTIVITY [J].

CEDAR, H .

CELL, 1988, 53 (01) :3-4

[5] The sounds of silence - histone deacetylation meets histone methylation [J].

Czermin, B ;

Imhof, A .

GENETICA, 2003, 117 (02) :159-164

[6] Regulation of heterochromatin by histone methylation and small RNAs [J].

Grewal, SIS ;

Rice, JC .

CURRENT OPINION IN CELL BIOLOGY, 2004, 16 (03) :230-238

[7]

He Hongpeng, 2003, Briefings in Functional Genomics & Proteomics, V2, P234, DOI 10.1093/bfgp/2.3.234

[8] BIOLOGICAL ASPECTS OF CYTOSINE METHYLATION IN EUKARYOTIC CELLS [J].

HERGERSBERG, M .

EXPERIENTIA, 1991, 47 (11-12) :1171-1185

[9] Time course of gene expression after plasmid DNA gene transfer to the liver [J].

Herweijer, H ;

Zhang, GF ;

Subbotin, VM ;

Budker, V ;

Williams, P ;

Wolff, JA .

JOURNAL OF GENE MEDICINE, 2001, 3 (03) :280-291

[10] Methylation of episomal plasmids as a barrier to transient gene expression via a synthetic delivery vector [J].

Hong, K ;

Sherley, J ;

Lauffenburger, DA .

BIOMOLECULAR ENGINEERING, 2001, 18 (04) :185-192

← 1 2 3 →