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Titolo:
Glowing zebrafish: Integration, transmission, and expression of a single luciferase transgene promoted by noncovalent DNA-nuclear transport peptide complexes
Autore:
Liang, MR; Alestrom, P; Collas, P;
Indirizzi:
Univ Oslo, Inst Med Biochem, N-0317 Oslo, Norway Univ Oslo Oslo Norway N-0317 Oslo, Inst Med Biochem, N-0317 Oslo, Norway Norwegian Coll Vet Med, Dept Biochem, Oslo, Norway Norwegian Coll Vet MedOslo Norway Vet Med, Dept Biochem, Oslo, Norway
Titolo Testata:
MOLECULAR REPRODUCTION AND DEVELOPMENT
fascicolo: 1, volume: 55, anno: 2000,
pagine: 8 - 13
SICI:
1040-452X(200001)55:1<8:GZITAE>2.0.ZU;2-P
Fonte:
ISI
Lingua:
ENG
Soggetto:
GERM-LINE TRANSMISSION; SV40 T-ANTIGEN; PLASMID DNA; LOCALIZATION SIGNAL; EMBRYO NUCLEI; SEQUENCES; METHYLATION; DIRECTS;
Keywords:
gene transfer; nuclear localization signal; peptide-DNA complex; Danio rerio;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
18
Recensione:
Indirizzi per estratti:
Indirizzo: Collas, P Univ Oslo, Inst Med Biochem, POB 1112, N-0317 Oslo, Norway Univ Oslo POB 1112 Oslo Norway N-0317 1112, N-0317 Oslo, Norway
Citazione:
M.R. Liang et al., "Glowing zebrafish: Integration, transmission, and expression of a single luciferase transgene promoted by noncovalent DNA-nuclear transport peptide complexes", MOL REPROD, 55(1), 2000, pp. 8-13

Abstract

The development of vehicles driving foreign DNA into the cell nucleus is essential for effective cellular gene transfer applications. We report that noncovalent binding of nuclear localization signal (NLS) peptides to plasmid DNA enhances nuclear uptake of the DNA and promotes germline integration,inheritance, and expression of a single copy of a luciferase reporter genein zebrafish. As few as 10 DNA-NLS complexes (0.06 fg plasmid DNA) cytoplasmically injected are sufficient to produce germline-transgenic zebrafish bearing a single copy of the transgene. This corresponds to a 10(5)-fold reduction in DNA concentration compared to commonly used procedures. Use of 10(3) or 10(4) DNA-NLS complexes augments the number of transgene integrations, which occur mostly within 1-4 distinct insertion sites in the genome. Insitu hybridization analyses and transmission studies show that transgene integration into the germline and somatic tissues is mosaic, and that the extent of mosaicism is negatively correlated with the amount of DNA-NLS injected. in addition, a larger proportion of zebrafish harboring a single copy of the transgene expresses luciferase, albeit at a 10-fold lower level thanthose containing numerous transgene insertions. The data demonstrate the potential use of nuclear targeting peptides noncovalently bound to Vector DNA to enhance the efficiency of biotechnological nonviral gene transfer applications. (C) 2000 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 29/03/20 alle ore 17:41:39