RRC ID 72156
Author Masuda T, Akita H, Harashima H.
Title Evaluation of nuclear transfer and transcription of plasmid DNA condensed with protamine by microinjection: the use of a nuclear transfer score.
Journal FEBS Lett
Abstract In the present study, the nuclear delivery of a green fluorescence protein (GFP)-encoding pDNA condensed by protamine was investigated in terms of trans-gene expression after cytoplasmic (E(cyt)) and nuclear (E(nuc)) microinjection. To compare the nuclear transfer process, a novel parameter; the nuclear transfer (NT) score was introduced. The E(cyt) value for protamine/pDNA particles increased in a charge ratio-dependent manner. The calculated NT score showed that this increase results from an enhancement in nuclear transfer efficiency, which was also quantitatively confirmed by a recently developed confocal image-assisted three-dimensionally integrated quantification (CIDIQ) method. Moreover, E(nuc) for protamine/pDNA particles was significantly higher than that for poly-L-lysine/pDNA particles, suggesting that pDNA, when condensed with protamine, is more accessible to intra-nuclear transcription. Collectively, protamine is an excellent DNA condenser, with bi-functional advantages: improvement in nuclear delivery and efficient intra-nuclear transcription.
Volume 579(10)
Pages 2143-8
Published 2005-4-11
DOI 10.1016/j.febslet.2005.02.071
PII S0014-5793(05)00319-4
PMID 15811332
MeSH Cell Nucleus* DNA / administration & dosage* DNA / chemistry Microinjections Plasmids* Protamines / chemistry* Transcription, Genetic*
Resource
Human and Animal Cells HeLa(RCB0007)