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Annexin A2 facilitates endocytic trafficking of antisense oligonucleotides
Chemically modified antisense oligonucleotides (ASOs) designed to mediate site-specific cleavage of RNA by RNase H1 are used as research tools and as therapeutics. ASOs modified with phosphorothioate (PS) linkages enter cells via endocytotic pathways. The mechanisms by which PS-ASOs are released fro...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5009748/ https://www.ncbi.nlm.nih.gov/pubmed/27378781 http://dx.doi.org/10.1093/nar/gkw595 |
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author | Wang, Shiyu Sun, Hong Tanowitz, Michael Liang, Xue-hai Crooke, Stanley T. |
author_facet | Wang, Shiyu Sun, Hong Tanowitz, Michael Liang, Xue-hai Crooke, Stanley T. |
author_sort | Wang, Shiyu |
collection | PubMed |
description | Chemically modified antisense oligonucleotides (ASOs) designed to mediate site-specific cleavage of RNA by RNase H1 are used as research tools and as therapeutics. ASOs modified with phosphorothioate (PS) linkages enter cells via endocytotic pathways. The mechanisms by which PS-ASOs are released from membrane-enclosed endocytotic organelles to reach target RNAs remain largely unknown. We recently found that annexin A2 (ANXA2) co-localizes with PS-ASOs in late endosomes (LEs) and enhances ASO activity. Here, we show that co-localization of ANXA2 with PS-ASO is not dependent on their direct interactions or mediated by ANXA2 partner protein S100A10. Instead, ANXA2 accompanies the transport of PS-ASOs to LEs, as ANXA2/PS-ASO co-localization was observed inside LEs. Although ANXA2 appears not to affect levels of PS-ASO internalization, ANXA2 reduction caused significant accumulation of ASOs in early endosomes (EEs) and reduced localization in LEs and decreased PS-ASO activity. Importantly, the kinetics of PS-ASO activity upon free uptake show that target mRNA reduction occurs at least 4 hrs after PS-ASOs exit from EEs and is coincident with release from LEs. Taken together, our results indicate that ANXA2 facilitates PS-ASO trafficking from early to late endosomes where it may also contribute to PS-ASO release. |
format | Online Article Text |
id | pubmed-5009748 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-50097482016-09-07 Annexin A2 facilitates endocytic trafficking of antisense oligonucleotides Wang, Shiyu Sun, Hong Tanowitz, Michael Liang, Xue-hai Crooke, Stanley T. Nucleic Acids Res Molecular Biology Chemically modified antisense oligonucleotides (ASOs) designed to mediate site-specific cleavage of RNA by RNase H1 are used as research tools and as therapeutics. ASOs modified with phosphorothioate (PS) linkages enter cells via endocytotic pathways. The mechanisms by which PS-ASOs are released from membrane-enclosed endocytotic organelles to reach target RNAs remain largely unknown. We recently found that annexin A2 (ANXA2) co-localizes with PS-ASOs in late endosomes (LEs) and enhances ASO activity. Here, we show that co-localization of ANXA2 with PS-ASO is not dependent on their direct interactions or mediated by ANXA2 partner protein S100A10. Instead, ANXA2 accompanies the transport of PS-ASOs to LEs, as ANXA2/PS-ASO co-localization was observed inside LEs. Although ANXA2 appears not to affect levels of PS-ASO internalization, ANXA2 reduction caused significant accumulation of ASOs in early endosomes (EEs) and reduced localization in LEs and decreased PS-ASO activity. Importantly, the kinetics of PS-ASO activity upon free uptake show that target mRNA reduction occurs at least 4 hrs after PS-ASOs exit from EEs and is coincident with release from LEs. Taken together, our results indicate that ANXA2 facilitates PS-ASO trafficking from early to late endosomes where it may also contribute to PS-ASO release. Oxford University Press 2016-09-06 2016-07-04 /pmc/articles/PMC5009748/ /pubmed/27378781 http://dx.doi.org/10.1093/nar/gkw595 Text en © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Molecular Biology Wang, Shiyu Sun, Hong Tanowitz, Michael Liang, Xue-hai Crooke, Stanley T. Annexin A2 facilitates endocytic trafficking of antisense oligonucleotides |
title | Annexin A2 facilitates endocytic trafficking of antisense oligonucleotides |
title_full | Annexin A2 facilitates endocytic trafficking of antisense oligonucleotides |
title_fullStr | Annexin A2 facilitates endocytic trafficking of antisense oligonucleotides |
title_full_unstemmed | Annexin A2 facilitates endocytic trafficking of antisense oligonucleotides |
title_short | Annexin A2 facilitates endocytic trafficking of antisense oligonucleotides |
title_sort | annexin a2 facilitates endocytic trafficking of antisense oligonucleotides |
topic | Molecular Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5009748/ https://www.ncbi.nlm.nih.gov/pubmed/27378781 http://dx.doi.org/10.1093/nar/gkw595 |
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