Integrase-mediated nonviral gene transfection with enhanced integration efficiency

Shinji Mizuarai, Masamichi Kamihira, Ken Ichi Nishijima, Shinji Iijima

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Retroviruses efficiently integrate their genome into the host chromosome. Two elements of the retrovirus genome are needed for the integration: long terminal repeats (LTRs) and integrase protein. We attempted to incorporate the retrovirus integration machinery in lipid vesicle-mediated gene transfection with the aim of achieving efficient stable transfection in a nonviral gene transfection system. A DNA fragment, in which a neomycin- resistant gene was flanked between partial LTR sequences derived from the Rous sarcoma virus (RSV), was constructed. This DNA fragment was transfected together with purified recombinant RSV integrase or integrase expression vectors by means of lipid vesicle-mediated gene transfection. The integrase- mediated transfection enhanced the stable transfection efficiency. The length and the end structure of the LTR sequences were important in achieving high efficiency. Under optimal conditions, the stable transfection efficiency showed a 16-fold improvement over that without integrase.

Original languageEnglish
Pages (from-to)461-467
Number of pages7
JournalJournal of Bioscience and Bioengineering
Volume88
Issue number5
DOIs
Publication statusPublished - Jan 1 1999
Externally publishedYes

Fingerprint

Integrases
Terminal Repeat Sequences
Transfection
Genes
Retroviridae
Rous sarcoma virus
Viruses
Lipids
DNA
Genome
Neomycin
Chromosomes
Machinery
Proteins

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Integrase-mediated nonviral gene transfection with enhanced integration efficiency. / Mizuarai, Shinji; Kamihira, Masamichi; Nishijima, Ken Ichi; Iijima, Shinji.

In: Journal of Bioscience and Bioengineering, Vol. 88, No. 5, 01.01.1999, p. 461-467.

Research output: Contribution to journalArticle

Mizuarai, Shinji ; Kamihira, Masamichi ; Nishijima, Ken Ichi ; Iijima, Shinji. / Integrase-mediated nonviral gene transfection with enhanced integration efficiency. In: Journal of Bioscience and Bioengineering. 1999 ; Vol. 88, No. 5. pp. 461-467.
@article{b3ae9feee3ff4d8abebd88a1af55fa46,
title = "Integrase-mediated nonviral gene transfection with enhanced integration efficiency",
abstract = "Retroviruses efficiently integrate their genome into the host chromosome. Two elements of the retrovirus genome are needed for the integration: long terminal repeats (LTRs) and integrase protein. We attempted to incorporate the retrovirus integration machinery in lipid vesicle-mediated gene transfection with the aim of achieving efficient stable transfection in a nonviral gene transfection system. A DNA fragment, in which a neomycin- resistant gene was flanked between partial LTR sequences derived from the Rous sarcoma virus (RSV), was constructed. This DNA fragment was transfected together with purified recombinant RSV integrase or integrase expression vectors by means of lipid vesicle-mediated gene transfection. The integrase- mediated transfection enhanced the stable transfection efficiency. The length and the end structure of the LTR sequences were important in achieving high efficiency. Under optimal conditions, the stable transfection efficiency showed a 16-fold improvement over that without integrase.",
author = "Shinji Mizuarai and Masamichi Kamihira and Nishijima, {Ken Ichi} and Shinji Iijima",
year = "1999",
month = "1",
day = "1",
doi = "10.1016/S1389-1723(00)87659-8",
language = "English",
volume = "88",
pages = "461--467",
journal = "Journal of Bioscience and Bioengineering",
issn = "1389-1723",
publisher = "The Society for Biotechnology, Japan",
number = "5",

}

TY - JOUR

T1 - Integrase-mediated nonviral gene transfection with enhanced integration efficiency

AU - Mizuarai, Shinji

AU - Kamihira, Masamichi

AU - Nishijima, Ken Ichi

AU - Iijima, Shinji

PY - 1999/1/1

Y1 - 1999/1/1

N2 - Retroviruses efficiently integrate their genome into the host chromosome. Two elements of the retrovirus genome are needed for the integration: long terminal repeats (LTRs) and integrase protein. We attempted to incorporate the retrovirus integration machinery in lipid vesicle-mediated gene transfection with the aim of achieving efficient stable transfection in a nonviral gene transfection system. A DNA fragment, in which a neomycin- resistant gene was flanked between partial LTR sequences derived from the Rous sarcoma virus (RSV), was constructed. This DNA fragment was transfected together with purified recombinant RSV integrase or integrase expression vectors by means of lipid vesicle-mediated gene transfection. The integrase- mediated transfection enhanced the stable transfection efficiency. The length and the end structure of the LTR sequences were important in achieving high efficiency. Under optimal conditions, the stable transfection efficiency showed a 16-fold improvement over that without integrase.

AB - Retroviruses efficiently integrate their genome into the host chromosome. Two elements of the retrovirus genome are needed for the integration: long terminal repeats (LTRs) and integrase protein. We attempted to incorporate the retrovirus integration machinery in lipid vesicle-mediated gene transfection with the aim of achieving efficient stable transfection in a nonviral gene transfection system. A DNA fragment, in which a neomycin- resistant gene was flanked between partial LTR sequences derived from the Rous sarcoma virus (RSV), was constructed. This DNA fragment was transfected together with purified recombinant RSV integrase or integrase expression vectors by means of lipid vesicle-mediated gene transfection. The integrase- mediated transfection enhanced the stable transfection efficiency. The length and the end structure of the LTR sequences were important in achieving high efficiency. Under optimal conditions, the stable transfection efficiency showed a 16-fold improvement over that without integrase.

UR - http://www.scopus.com/inward/record.url?scp=0033404828&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033404828&partnerID=8YFLogxK

U2 - 10.1016/S1389-1723(00)87659-8

DO - 10.1016/S1389-1723(00)87659-8

M3 - Article

C2 - 16232645

AN - SCOPUS:0033404828

VL - 88

SP - 461

EP - 467

JO - Journal of Bioscience and Bioengineering

JF - Journal of Bioscience and Bioengineering

SN - 1389-1723

IS - 5

ER -