Efficient DNA transfection in neuronal and astrocytic cell lines

Chandramallika Ghosh, Weihong Song, Debomoy K. Lahiri

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

We have studied different parameters for efficient DNA transfection in various cell types and with different size of the promoter. Here we report that the optimum condition for DNA transfection by electroporation is 350 V/960 μF for PC12, 450V/960 μF C6 cells, and 250 V/500 μF for COS-1 cells. For the human neuroblastoma (SK-N-SH) cells the optimum condition for DNA transfection is by the calcium phosphate method. In promoter mapping studies, a serial deletion approach is commonly used. To optimize transfection we have selected three DNA constructs that varied in size from 4.5 to 12.4 kilobases (kb). We measured the promoter activity of these constructs under conditions of 'equal amount', 'equimolar', and 'equimolar plus carrier DNA to make it equal amount'. We recommend that for comparative purpose, transfection should be carried out under 'equimolar condition' without a need to adjust the total amount of DNA by carrier DNA. Taken together, our results suggest that efficient methods for DNA transfection are important to study gene regulation by devising better ways to deliver DNA into the mammalian cells.

Original languageEnglish (US)
Pages (from-to)113-121
Number of pages9
JournalMolecular Biology Reports
Volume27
Issue number2
DOIs
StatePublished - Nov 18 2000

Fingerprint

Transfection
Cells
Cell Line
DNA
Electroporation
COS Cells
Neuroblastoma
Gene expression
Genes

Keywords

  • Calcium phosphate
  • Electroporation
  • Neuroblastoma cells
  • PC12 cells
  • Promoter
  • Reporter gene
  • β-amyloid precursor protein

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Efficient DNA transfection in neuronal and astrocytic cell lines. / Ghosh, Chandramallika; Song, Weihong; Lahiri, Debomoy K.

In: Molecular Biology Reports, Vol. 27, No. 2, 18.11.2000, p. 113-121.

Research output: Contribution to journalArticle

Ghosh, Chandramallika ; Song, Weihong ; Lahiri, Debomoy K. / Efficient DNA transfection in neuronal and astrocytic cell lines. In: Molecular Biology Reports. 2000 ; Vol. 27, No. 2. pp. 113-121.
@article{057546250be84ddd998cbed1f60d6f0e,
title = "Efficient DNA transfection in neuronal and astrocytic cell lines",
abstract = "We have studied different parameters for efficient DNA transfection in various cell types and with different size of the promoter. Here we report that the optimum condition for DNA transfection by electroporation is 350 V/960 μF for PC12, 450V/960 μF C6 cells, and 250 V/500 μF for COS-1 cells. For the human neuroblastoma (SK-N-SH) cells the optimum condition for DNA transfection is by the calcium phosphate method. In promoter mapping studies, a serial deletion approach is commonly used. To optimize transfection we have selected three DNA constructs that varied in size from 4.5 to 12.4 kilobases (kb). We measured the promoter activity of these constructs under conditions of 'equal amount', 'equimolar', and 'equimolar plus carrier DNA to make it equal amount'. We recommend that for comparative purpose, transfection should be carried out under 'equimolar condition' without a need to adjust the total amount of DNA by carrier DNA. Taken together, our results suggest that efficient methods for DNA transfection are important to study gene regulation by devising better ways to deliver DNA into the mammalian cells.",
keywords = "Calcium phosphate, Electroporation, Neuroblastoma cells, PC12 cells, Promoter, Reporter gene, β-amyloid precursor protein",
author = "Chandramallika Ghosh and Weihong Song and Lahiri, {Debomoy K.}",
year = "2000",
month = "11",
day = "18",
doi = "10.1023/A:1007173906990",
language = "English (US)",
volume = "27",
pages = "113--121",
journal = "Molecular Biology Reports",
issn = "0301-4851",
publisher = "Springer Netherlands",
number = "2",

}

TY - JOUR

T1 - Efficient DNA transfection in neuronal and astrocytic cell lines

AU - Ghosh, Chandramallika

AU - Song, Weihong

AU - Lahiri, Debomoy K.

PY - 2000/11/18

Y1 - 2000/11/18

N2 - We have studied different parameters for efficient DNA transfection in various cell types and with different size of the promoter. Here we report that the optimum condition for DNA transfection by electroporation is 350 V/960 μF for PC12, 450V/960 μF C6 cells, and 250 V/500 μF for COS-1 cells. For the human neuroblastoma (SK-N-SH) cells the optimum condition for DNA transfection is by the calcium phosphate method. In promoter mapping studies, a serial deletion approach is commonly used. To optimize transfection we have selected three DNA constructs that varied in size from 4.5 to 12.4 kilobases (kb). We measured the promoter activity of these constructs under conditions of 'equal amount', 'equimolar', and 'equimolar plus carrier DNA to make it equal amount'. We recommend that for comparative purpose, transfection should be carried out under 'equimolar condition' without a need to adjust the total amount of DNA by carrier DNA. Taken together, our results suggest that efficient methods for DNA transfection are important to study gene regulation by devising better ways to deliver DNA into the mammalian cells.

AB - We have studied different parameters for efficient DNA transfection in various cell types and with different size of the promoter. Here we report that the optimum condition for DNA transfection by electroporation is 350 V/960 μF for PC12, 450V/960 μF C6 cells, and 250 V/500 μF for COS-1 cells. For the human neuroblastoma (SK-N-SH) cells the optimum condition for DNA transfection is by the calcium phosphate method. In promoter mapping studies, a serial deletion approach is commonly used. To optimize transfection we have selected three DNA constructs that varied in size from 4.5 to 12.4 kilobases (kb). We measured the promoter activity of these constructs under conditions of 'equal amount', 'equimolar', and 'equimolar plus carrier DNA to make it equal amount'. We recommend that for comparative purpose, transfection should be carried out under 'equimolar condition' without a need to adjust the total amount of DNA by carrier DNA. Taken together, our results suggest that efficient methods for DNA transfection are important to study gene regulation by devising better ways to deliver DNA into the mammalian cells.

KW - Calcium phosphate

KW - Electroporation

KW - Neuroblastoma cells

KW - PC12 cells

KW - Promoter

KW - Reporter gene

KW - β-amyloid precursor protein

UR - http://www.scopus.com/inward/record.url?scp=0033755065&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033755065&partnerID=8YFLogxK

U2 - 10.1023/A:1007173906990

DO - 10.1023/A:1007173906990

M3 - Article

C2 - 11092558

AN - SCOPUS:0033755065

VL - 27

SP - 113

EP - 121

JO - Molecular Biology Reports

JF - Molecular Biology Reports

SN - 0301-4851

IS - 2

ER -