Abstract
A 3,-[[2-[N-(3-aminopropyl)-N-(2-hydroxyethyl)amino]ethylphosphoryl]oligodeoxyribonucleoside methylphosphonate 12-mer was synthesized using the Aha-CPG solid support [Thaden, J. and Miller, P. S. (1993) Bioconjugate Chem., companion paper in this issue]. The oligomer was conjugated at the 3′ primary aliphatic amine with tetramethylrhodamine 5-isothiocyanate. The rhodamine linker/spacer was stable in 10% fetal calf serum. After enzymatic phosphorylation, the molecule was conjugated at the 5′ phosphate with 4′-[N-(2-aminoethyl)aminomethyl]-4,5′,8-trimethylpsoralen [(ae(AMT)]. The rhodamine/psoralen doubly-conjugated oligomer formed photoadducts with complementary singlestranded DNA and RNA oligonucleotides when irradiated with long-wavelength ultraviolet light. The efficiency of UV crosslinking slightly exceeded that of a colinear, psoralen-derivatized oligonucleoside methylphosphonate, and exhibited relationships with UV fluence and temperature that are characteristic for psoralen-conjugated methylphosphonates. The 1:1 complex formed with the oligodeoxyribonucleotide target could be detected by its red fluorescence. Mouse L949 cells grown in the presence of the double conjugate were shown by means of computer-assisted epifluorescence microscopy to have internalized it. There was an accumulation of intensely fluorescent points and spots in a juxtanuclear region of the cytoplasm, and a faint, diffuse signal in the entire cell area.
Original language | English (US) |
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Pages (from-to) | 386-394 |
Number of pages | 9 |
Journal | Bioconjugate Chemistry |
Volume | 4 |
Issue number | 5 |
DOIs | |
State | Published - Sep 1 1993 |
Externally published | Yes |
ASJC Scopus subject areas
- Biotechnology
- Bioengineering
- Biomedical Engineering
- Pharmacology
- Pharmaceutical Science
- Organic Chemistry