Technical Reference #375

375.

Direct Observation Of Specific Messenger RNA In A Single Living Cell Under A Fluorescence Microscope Akihiko Tsuji; Hiroyuki Koshimoto; Yoshihiro Sato; Masahiko Hirano; Yukari Sei-Iida; Satoshi Kondo; and Kaname Ishibashi, Laboratory of Molecular Biophotonics, Biophysical Journal, 78(375), (2000)
Link To Paper

Abstract:
We observed the expression of human c-fos mRNA in a living transfected Cos7 cell under a fluorescencemicroscope by detecting hybrid formed with two fluorescently labeled oligodeoxynucleotides (oligoDNAs) and c-fos mRNAin the cytoplasm. Two fluorescent

Keywords:
mRNA; oligodeoxynucleotides (oligoDNAs); fluorescence resonance energy transfer (FRET); posttranscriptional genetic control

Materials & Methods:
The pSPT-cfos was treated with EcoRI to cut off c-fos DNA (2.1 kb) and the c-fos DNA fragment was inserted into the EcoRI site on pME18S expression vector driven by SRa promoter (pME18S-cfos). Cos7 cells were cultured with Dulbecco’s modified Eagle’s

Microscopic Technique
Fluorescence Microscopy

Cell Type(s)
COS-7