Evaluation of nuclear transfer and transcription of plasmid DNA condensed with protamine by microinjection: The use of a nuclear transfer score

被引：69

作者：

Masuda, T ^{[1
]}

Akita, H ^{[1
]}

Harashima, H ^{[1
]}

机构：

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

来源：

FEBS LETTERS | 2005年 / 579卷 / 10期

关键词：

protamine; microinjection; nuclear delivery; gene delivery; plasmid DNA;

D O I：

10.1016/j.febslet.2005.02.071

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：

In the present study, the nuclear delivery of a green fluorescence protein (GFP)-encoding pDNA condensed by protamine was investigated in terms of trans-gene expression after cytoplasmic (E(cyt)) and nuclear (E(nuc)) microinjection. To compare the nuclear transfer process, a novel parameter; the nuclear transfer (NT) score was introduced. The E(cyt) value for protamine/pDNA particles increased in a charge ratio-dependent manner. The calculated NT score showed that this increase results from an enhancement in nuclear transfer efficiency, which was also quantitatively confirmed by a recently developed confocal image-assisted three-dimensionally integrated quantification (CIDIQ) method. Moreover, E(nuc) for protamine/pDNA particles was significantly higher than that for poly-L-lysine/pDNA particles. suggesting that pDNA, when condensed with protamine, is more accessible to intra-nuclear transcription. Collectively, protamine is an excellent DNA condenser, with bi-functional advantages: improvement in nuclear delivery and efficient intra-nuclear transcription. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

引用

页码：2143 / 2148

页数：6

共 38 条

[1] Quantitative three-dimensional analysis of the intracellular trafficking of plasmid DNA transfected by a nonviral gene delivery system using confocal laser scanning microscopy [J].

Akita, H ;

Ito, R ;

Khalil, IA ;

Futaki, S ;

Harashima, H .

MOLECULAR THERAPY, 2004, 9 (03) :443-451

[2] A VERSATILE VECTOR FOR GENE AND OLIGONUCLEOTIDE TRANSFER INTO CELLS IN CULTURE AND IN-VIVO - POLYETHYLENIMINE [J].

BOUSSIF, O ;

LEZOUALCH, F ;

ZANTA, MA ;

MERGNY, MD ;

SCHERMAN, D ;

DEMENEIX, B ;

BEHR, JP .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (16) :7297-7301

[3] Roles of NPM2 in chromatin and nucleolar organization in oocytes and embryos [J].

Burns, KH ;

Viveiros, MM ;

Ren, YS ;

Wang, P ;

DeMayo, FJ ;

Frail, DE ;

Eppig, JJ ;

Matzuk, MM .

SCIENCE, 2003, 300 (5619) :633-636

[4] LIPIDS OF MITOCHONDRIA [J].

DAUM, G .

BIOCHIMICA ET BIOPHYSICA ACTA, 1985, 822 (01) :1-42

[5] THE ROLE OF DIOLEOYL PHOSPHATIDYLETHANOLAMINE IN CATIONIC LIPOSOME-MEDIATED GENE-TRANSFER [J].

FARHOOD, H ;

SERBINA, N ;

HUANG, L .

BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 1995, 1235 (02) :289-295

[6] Intracellular control of gene trafficking using liposomes as drug carriers [J].

Harashima, H ;

Shinohara, Y ;

Kiwada, H .

EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES, 2001, 13 (01) :85-89

[7] Ternary complex formation between DNA-adenovirus core protein VII and TAF-Iβ/SET, an acidic molecular chaperone [J].

Haruki, H ;

Gyurcsik, B ;

Okuwaki, M ;

Nagata, K .

FEBS LETTERS, 2003, 555 (03) :521-527

[8] The role of helper lipids in cationic liposome-mediated gene transfer [J].

Hui, SW ;

Langner, M ;

Zhao, YL ;

Ross, P ;

Hurley, E ;

Chan, K .

BIOPHYSICAL JOURNAL, 1996, 71 (02) :590-599

[9] Visualization of transcription-dependent association of imprinted genes with the nuclear matrix [J].

Kagotani, K ;

Nabeshima, H ;

Kohda, A ;

Nakao, M ;

Taguchi, H ;

Okumura, K .

EXPERIMENTAL CELL RESEARCH, 2002, 274 (02) :189-196

[10] Transferrin-modified liposomes equipped with a pH-sensitive fusogenic peptide: An artificial viral-like delivery system [J].

Kakudo, T ;

Chaki, S ;

Futaki, S ;

Nakase, I ;

Akaji, K ;

Kawakami, T ;

Maruyama, K ;

Kamiya, H ;

Harashima, H .

BIOCHEMISTRY, 2004, 43 (19) :5618-5628

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