Sub-cellular temporal and spatial distribution of electrotransferred LNA/DNA oligomer
نویسندگان
چکیده
Low biological activity and inefficient targeted delivery in vivo have hindered RNA interference (RNAi)-based therapy from realising its full clinical potential. To overcome these hurdles, progresses have been made to develop new technologies optimizing oligonucleotides chemistry on one hand and achieving its effective delivery on the other hand. In this report, we achieved, by using the electropulsation technique (EP), efficient cellular delivery of chemically-modified oligonucleotide: The locked nucleic acid (LNA)/DNA oligomer. We used single cell level confocal fluorescence microscopy to follow the spatial and temporal distribution of electrotransferred cyanine 5 (Cy5)-labeled LNA/DNA oligomer. We observed that EP allowed LNA/DNA oligomer cellular uptake providing the oligomer a rapid access to the cytoplasm of HeLa cells. Within a few minutes after electrotransfer, Cy5-LNA/DNA oligomers shuttle from cytoplasm to nucleus whereas in absence of pulses application, Cy5-LNA/DNA oligomers were not detected. We then observed a redistribution of the Cy5 fluorescence that accumulated over time into cytoplasmic organelles. To go further and to identify these compartments, we used the HeLa GFP-Rab7 cell line to visualise late endosomes, and lysosomal or mitochondrial specific markers. Our results showed that the EP technique allowed direct entry into the cytoplasm of the Cy5-LNA/DNA oligomer bypassing the endocytosic pathway. However, in absence of pulses application, Cy5-LNA/DNA oligomer were able to enter cells through the endocytosic pathway. We demonstrated that EP is an efficient technique for LNA-based oligonucleotides delivery offering strong advantages by avoiding the endolysosomal compartmentalization, giving a rapid and free access to the cytoplasm and the nucleus where they can find their targets.
منابع مشابه
Temporal and spatial distribution of extreme precipitation indices over the lake Urmia Basin, Iran
متن کامل
Spatio-temporal patterns of crab fisheries in the main bays of Guangdong Province, China
Using a semi-balloon otter trawl, crab fisheries in the main bays of Guangdong Province, China, were carried out seasonally . A total of 70 species were found, all belonging to the South China Sea Faunal sub region in the tropical India-West-Pacific Faunal Region. The clustering and nMDS ordination analysis revealed the existence of three groups. Group 1 included Hailing Bay and four bays to ...
متن کاملTemporal and spatial distribution pattern of Bullacta exarata in a tidal flat at south shore of Hangzhou Bay, China
The distribution pattern of Bullacta exarata was studied in different seasons of 2004 at south shore of Hangzhou Bay, China. We found that the distribution pattern of B. exarata was aggregated in each season by Taylor's power regression and Iwao's plot regresses methods (P < 0.001). Based on two-way ANOVA analysis, the results indicated that the densities were significantly affected by the fact...
متن کاملTemporal and spatial distribution pattern of Bullacta exarata in a tidal flat at south shore of Hangzhou Bay, China
The distribution pattern of Bullacta exarata was studied in different seasons of 2004 at south shore of Hangzhou Bay, China. We found that the distribution pattern of B. exarata was aggregated in each season by Taylor's power regression and Iwao's plot regresses methods (P < 0.001). Based on two-way ANOVA analysis, the results indicated that the densities were significantly affected by the fact...
متن کاملModeling and Spatio-Temporal Analysis of the Distribution of O3 in Tehran City Based on Neural Network and Spatial Analysis in GIS Environment
Air pollution is one of the most problems that people are facing today in metropolitan areas. Suspended particulates, carbon monoxide, sulfur dioxide, ozone and nitrogen dioxide are the five major pollutants of air that pose many problems to human health. The goal of this study is to propose a spatial approach for estimation and analyzing the spatial and temporal distribution of ozone based on ...
متن کامل