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DNA uptake and repair enzyme access to transfected DNA is under reported by gene expression.

Abstract：

:Gene expression is the typical biological end point of interest following transfection. However, transcription may not accurately assess DNA uptake, or the ability of transfected DNA to be acted on by other enzymatic pathways. We have compared DNA uptake to gene expression and the unrelated enzymatic process of DNA double strand break (DSB) repair. Transfection efficiency (at limiting DNA concentration) was assessed as a function of DNA uptake and gene expression in the DSB repair proficient WI38VA13 and MO59K cell lines and the DSB repair defective cell line MO59J, by comparing eGFP expression from the pHygEGFP expression vector with uptake of rhodamine labeled linear pSP189 plasmid (3:1). Repair proficient cells expressed eGFP most efficiently, but never approached DNA uptake levels (>or=90%). Although transfected DNAs were stable in repair proficient cells and degraded in MO59J cells, most cells did not express eGFP, but in the repair proficient cells linear DNA did undergo DSB repair.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Pastwa E,Lubner EM,Mezhevaya K,Neumann RD,Winters TA

doi

10.1016/s0006-291x(03)00972-0

subject

Has Abstract

pub_date

2003-06-27 00:00:00

pages

421-9

issue

2

eissn

0006-291X

issn

1090-2104

pii

S0006291X03009720

journal_volume

306

pub_type

杂志文章

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