PMID- 21641986
OWN - NLM
STAT- MEDLINE
DCOM- 20111123
LR  - 20110711
IS  - 1873-3476 (Electronic)
IS  - 0378-5173 (Linking)
VI  - 415
IP  - 1-2
DP  - 2011 Aug 30
TI  - Dual imaging of mRNA and protein production: an investigation of the mechanism of 
      heterogeneity in cationic lipid-mediated transgene expression.
PG  - 218-20
LID - 10.1016/j.ijpharm.2011.05.051 [doi]
AB  - Heterogeneity of transgene expression is a severe disadvantage in the use of 
      cationic lipid-mediated gene vectors. We previously demonstrated that 
      heterogeneity of the post-nuclear delivery process, as well as intracellular 
      trafficking (i.e. nuclear delivery) is a major determinant in the overall 
      heterogeneity in gene expression, when plasmid DNA (pDNA) is transfected to HeLa 
      cells using a lipoplex. In this study, we explored the mechanism underlying this 
      heterogeneity in a post-nuclear transport process by the dual imaging of mRNA and 
      its encoded protein (histone H2B-tagged mTFP1; mTFP1-H2B) in a single cell. To 
      establish a highly sensitive imaging system for mRNA, we used fluorescence in 
      situ hybridization (FISH) combined with tyramide signal amplification (TSA) and a 
      semiconductor quantum dot (QD) probe. The mRNA expression and protein production 
      were quantified by counting the total pixel intensity in the region of interest 
      (r.o.i.) surrounding single cells. As a result, the correlation was poor in a 
      scattered plot of mRNA expression versus protein production in individual cells. 
      These findings demonstrate that cell-to-cell differences in the translation 
      process are also a key factor in heterogeneous gene expression.
CI  - Copyright (c) 2011 Elsevier B.V. All rights reserved.
FAU - Akita, Hidetaka
AU  - Akita H
AD  - Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo, 
      Hokkaido 060-0812, Japan. akita@pharm.hokudai.ac.jp
FAU - Umetsu, Yuko
AU  - Umetsu Y
FAU - Kurihara, Dai
AU  - Kurihara D
FAU - Harashima, Hideyoshi
AU  - Harashima H
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110527
PL  - Netherlands
TA  - Int J Pharm
JT  - International journal of pharmaceutics
JID - 7804127
RN  - 0 (Cations)
RN  - 0 (Histones)
RN  - 0 (Lipids)
RN  - 0 (Luminescent Proteins)
RN  - 0 (Proteins)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Cations
MH  - *Gene Expression
MH  - HeLa Cells
MH  - Histones/genetics
MH  - Humans
MH  - Lipids/*chemistry
MH  - Luminescent Proteins
MH  - Molecular Imaging/*methods
MH  - Protein Biosynthesis
MH  - Protein Transport
MH  - Proteins/*genetics
MH  - Quantum Dots
MH  - RNA, Messenger/*genetics
MH  - Transfection
MH  - *Transgenes
EDAT- 2011/06/07 06:00
MHDA- 2011/12/13 00:00
CRDT- 2011/06/07 06:00
PHST- 2011/03/09 00:00 [received]
PHST- 2011/05/02 00:00 [revised]
PHST- 2011/05/19 00:00 [accepted]
PHST- 2011/06/07 06:00 [entrez]
PHST- 2011/06/07 06:00 [pubmed]
PHST- 2011/12/13 00:00 [medline]
AID - S0378-5173(11)00499-6 [pii]
AID - 10.1016/j.ijpharm.2011.05.051 [doi]
PST - ppublish
SO  - Int J Pharm. 2011 Aug 30;415(1-2):218-20. doi: 10.1016/j.ijpharm.2011.05.051. 
      Epub 2011 May 27.