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Inhibition of inositol uptake in astrocytes by antisense oligonucleotides delivered by pH‐sensitive liposomes

Author:
Lubrich, Beate, van Calker, Dietrich, Peschka‐Süss, Regine
Source:
European journal of biochemistry 2000 v.267 no.8 pp. 2432-2438
ISSN:
0014-2956
Subject:
astrocytes, encapsulation, messenger RNA, myo-inositol, oligonucleotides, reverse transcriptase polymerase chain reaction
Abstract:
An oligonucleotide of 20 bases, complementary to a region of the sodium/myo‐inositol 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 liposome‐encapsulated material. Delivery of liposome‐encapsulated material (marker or oligonucleotides) into astrocytoma cells and primary astrocyte cultures was more effective with pH‐sensitive liposomes [dioleoylphosphatidylethanolamine (DOPE)/cholesteryl hemisuccinate (CHEMS)] than with non‐pH‐sensitive liposomes (soy lecithin) or free material in solution. Antisense activity was evaluated by determination of myo‐inositol uptake and detection of SMIT transcripts by RT‐PCR. Encapsulation of oligonucleotides in pH‐sensitive liposomes increased the inhibition of inositol uptake at least 50‐fold compared with application of free oligonucleotides in solution.
Agid:
206149