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Inhibition of inositol uptake in astrocytes by antisense oligonucleotides delivered by pHâsensitive liposomes
- Lubrich, Beate, van Calker, Dietrich, PeschkaâSÃ¼ss, Regine
- European journal of biochemistry 2000 v.267 no.8 pp. 2432-2438
- astrocytes, encapsulation, messenger RNA, myo-inositol, oligonucleotides, reverse transcriptase polymerase chain reaction
- An oligonucleotide of 20 bases, complementary to a region of the sodium/myoâinositol cotransporter (SMIT) mRNA, was used to investigate the uptake efficiency and activity of transferred antisense oligonucleotides with regard to substrate uptake. We compared the efficiency of oligonucleotide delivery after application of either free or liposomeâencapsulated material. Delivery of liposomeâencapsulated material (marker or oligonucleotides) into astrocytoma cells and primary astrocyte cultures was more effective with pHâsensitive liposomes [dioleoylphosphatidylethanolamine (DOPE)/cholesteryl hemisuccinate (CHEMS)] than with nonâpHâsensitive liposomes (soy lecithin) or free material in solution. Antisense activity was evaluated by determination of myoâinositol uptake and detection of SMIT transcripts by RTâPCR. Encapsulation of oligonucleotides in pHâsensitive liposomes increased the inhibition of inositol uptake at least 50âfold compared with application of free oligonucleotides in solution.