Intracellular Uptake of Modified Oligonucleotide Studied by Two Fluorescence Techniques

Abstract

Cellular uptake and intracellular distribution of synthetic modified antisense oligonucleotide with B16 melanoma cell line was studied employing cationic polyene antibiotic, amphotericin B 3-dimethylaminopropyl amide, as a carrier vector. The antisense oligonucleotide - dT15 oligomer analogue containing linkages 3'-O-P-CH2-O-5' - was labeled with TMR marker.

Strong preference staining of the cell nucleus was found. Fluorescence lifetime measurements from the intracellular environment yielded two spectral components of 4.9ns and 1.4ns lifetime, respectively.