DNA transfection of mouse lymphoid cells by the combination of DEAE-dextran-mediated DNA uptake and osmotic shock procedure

Toshiyuki Takai; Hitoshi Ohmori

doi:10.1016/0167-4781(90)90029-2

DNA transfection of mouse lymphoid cells by the combination of DEAE-dextran-mediated DNA uptake and osmotic shock procedure

Toshiyuki Takai, Hitoshi Ohmori

Research output: Contribution to journal › Article › peer-review

13 Citations (Scopus)

Abstract

Several mouse lymphoid cell lines were efficiently transfected with plasmid DNA by a novel method combining DEAE-dextran-mediated DNA uptake and osmotic shock procedure. The cells were first incubated with DNA-DEAE-dextran complex, treated with hypertonic Tris-HCl buffer containing 0.5 M sucrose and 10% poly(ethylene glycol), and then exposed to hypotonic RPMI 1640 medium. This transfection protocol exhibited maximal frequencies of 0.3% and 3·10^-5 for transient gene expression and stable transformation in P3-NSI/1-Ag4-1 cells, respectively.

Original language	English
Pages (from-to)	105-109
Number of pages	5
Journal	BBA - Gene Structure and Expression
Volume	1048
Issue number	1
DOIs	https://doi.org/10.1016/0167-4781(90)90029-2
Publication status	Published - 1990 Jan 30
Externally published	Yes

Keywords

(Mouse lymphoid cell)
DEAE-dextran
DNA transfection
Osmotic shock

ASJC Scopus subject areas

Structural Biology
Biophysics
Biochemistry
Genetics

Access to Document

10.1016/0167-4781(90)90029-2

Cite this

@article{5c93be35e40f497b8ed44349b17a9ef0,

title = "DNA transfection of mouse lymphoid cells by the combination of DEAE-dextran-mediated DNA uptake and osmotic shock procedure",

abstract = "Several mouse lymphoid cell lines were efficiently transfected with plasmid DNA by a novel method combining DEAE-dextran-mediated DNA uptake and osmotic shock procedure. The cells were first incubated with DNA-DEAE-dextran complex, treated with hypertonic Tris-HCl buffer containing 0.5 M sucrose and 10% poly(ethylene glycol), and then exposed to hypotonic RPMI 1640 medium. This transfection protocol exhibited maximal frequencies of 0.3% and 3·10-5 for transient gene expression and stable transformation in P3-NSI/1-Ag4-1 cells, respectively.",

keywords = "(Mouse lymphoid cell), DEAE-dextran, DNA transfection, Osmotic shock",

author = "Toshiyuki Takai and Hitoshi Ohmori",

note = "Funding Information: This work was supported in part by research grants from the Ministry of Education, Science and Culture of Japan.",

year = "1990",

month = jan,

day = "30",

doi = "10.1016/0167-4781(90)90029-2",

language = "English",

volume = "1048",

pages = "105--109",

journal = "Biochimica et Biophysica Acta - Gene Structure and Expression",

issn = "0167-4781",

publisher = "Elsevier BV",

number = "1",

}

TY - JOUR

T1 - DNA transfection of mouse lymphoid cells by the combination of DEAE-dextran-mediated DNA uptake and osmotic shock procedure

AU - Takai, Toshiyuki

AU - Ohmori, Hitoshi

N1 - Funding Information: This work was supported in part by research grants from the Ministry of Education, Science and Culture of Japan.

PY - 1990/1/30

Y1 - 1990/1/30

N2 - Several mouse lymphoid cell lines were efficiently transfected with plasmid DNA by a novel method combining DEAE-dextran-mediated DNA uptake and osmotic shock procedure. The cells were first incubated with DNA-DEAE-dextran complex, treated with hypertonic Tris-HCl buffer containing 0.5 M sucrose and 10% poly(ethylene glycol), and then exposed to hypotonic RPMI 1640 medium. This transfection protocol exhibited maximal frequencies of 0.3% and 3·10-5 for transient gene expression and stable transformation in P3-NSI/1-Ag4-1 cells, respectively.

AB - Several mouse lymphoid cell lines were efficiently transfected with plasmid DNA by a novel method combining DEAE-dextran-mediated DNA uptake and osmotic shock procedure. The cells were first incubated with DNA-DEAE-dextran complex, treated with hypertonic Tris-HCl buffer containing 0.5 M sucrose and 10% poly(ethylene glycol), and then exposed to hypotonic RPMI 1640 medium. This transfection protocol exhibited maximal frequencies of 0.3% and 3·10-5 for transient gene expression and stable transformation in P3-NSI/1-Ag4-1 cells, respectively.

KW - (Mouse lymphoid cell)

KW - DEAE-dextran

KW - DNA transfection

KW - Osmotic shock

UR - http://www.scopus.com/inward/record.url?scp=0025177278&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025177278&partnerID=8YFLogxK

U2 - 10.1016/0167-4781(90)90029-2

DO - 10.1016/0167-4781(90)90029-2

M3 - Article

C2 - 2297528

AN - SCOPUS:0025177278

SN - 0167-4781

VL - 1048

SP - 105

EP - 109

JO - Biochimica et Biophysica Acta - Gene Structure and Expression

JF - Biochimica et Biophysica Acta - Gene Structure and Expression

IS - 1

ER -

DNA transfection of mouse lymphoid cells by the combination of DEAE-dextran-mediated DNA uptake and osmotic shock procedure

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this