Remnant cationic dendrimers block RNA migration in electrophoresis after monophasic lysis
Cationic dendrimers such as poly(amidoamine) (PAMAM) and poly(propyleneimine) (PPI) have attractive characteristics for the delivery of nucleic acid and various biomedical applications. Most studies have focused on cationic dendrimer-based intracellular delivery, and very few studies have focused on...
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Veröffentlicht in: | Journal of biotechnology 2007-05, Vol.129 (3), p.383-390 |
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description | Cationic dendrimers such as poly(amidoamine) (PAMAM) and poly(propyleneimine) (PPI) have attractive characteristics for the delivery of nucleic acid and various biomedical applications. Most studies have focused on cationic dendrimer-based intracellular delivery, and very few studies have focused on the non-specific interaction of remnant cationic dendrimers with total RNA after isolation directly from cells in vitro. We examined RNA isolation using the common method of monophasic lysis from human macrophage-like cells (U937) and mouse fibroblast cells (NIH/3T3) that had been exposed to dendrimers and DNA/dendrimer complexes using gel electrophoresis. We found that PAMAM and PPI dendrimers strongly altered the mobility of RNA in the gels. In addition, the extent of dendrimer-induced alteration in RNA mobility was directly dendrimer-generation-dependent: the alteration was greater with higher-generation dendrimers. We also found that DNA/dendrimer complexes at higher dendrimer to DNA ratios interacted with RNA after isolation while gene expression was maintained. The interactions between RNA and remnant dendrimers after isolation were caused by electrostatic bindings, and we recovered total RNA using high ionic strength solvents (2M NaCl solution) to disrupt the electrostatic forces binding dendrimers to RNA. Because RNA isolation is routinely used for biological applications, such dendrimer-induced alteration in RNA mobility should be accounted for in the further processing of RNA-related applications. |
doi_str_mv | 10.1016/j.jbiotec.2007.01.030 |
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Most studies have focused on cationic dendrimer-based intracellular delivery, and very few studies have focused on the non-specific interaction of remnant cationic dendrimers with total RNA after isolation directly from cells in vitro. We examined RNA isolation using the common method of monophasic lysis from human macrophage-like cells (U937) and mouse fibroblast cells (NIH/3T3) that had been exposed to dendrimers and DNA/dendrimer complexes using gel electrophoresis. We found that PAMAM and PPI dendrimers strongly altered the mobility of RNA in the gels. In addition, the extent of dendrimer-induced alteration in RNA mobility was directly dendrimer-generation-dependent: the alteration was greater with higher-generation dendrimers. We also found that DNA/dendrimer complexes at higher dendrimer to DNA ratios interacted with RNA after isolation while gene expression was maintained. The interactions between RNA and remnant dendrimers after isolation were caused by electrostatic bindings, and we recovered total RNA using high ionic strength solvents (2M NaCl solution) to disrupt the electrostatic forces binding dendrimers to RNA. 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Most studies have focused on cationic dendrimer-based intracellular delivery, and very few studies have focused on the non-specific interaction of remnant cationic dendrimers with total RNA after isolation directly from cells in vitro. We examined RNA isolation using the common method of monophasic lysis from human macrophage-like cells (U937) and mouse fibroblast cells (NIH/3T3) that had been exposed to dendrimers and DNA/dendrimer complexes using gel electrophoresis. We found that PAMAM and PPI dendrimers strongly altered the mobility of RNA in the gels. In addition, the extent of dendrimer-induced alteration in RNA mobility was directly dendrimer-generation-dependent: the alteration was greater with higher-generation dendrimers. We also found that DNA/dendrimer complexes at higher dendrimer to DNA ratios interacted with RNA after isolation while gene expression was maintained. The interactions between RNA and remnant dendrimers after isolation were caused by electrostatic bindings, and we recovered total RNA using high ionic strength solvents (2M NaCl solution) to disrupt the electrostatic forces binding dendrimers to RNA. Because RNA isolation is routinely used for biological applications, such dendrimer-induced alteration in RNA mobility should be accounted for in the further processing of RNA-related applications.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cationic dendrimer</subject><subject>Dendrimers - metabolism</subject><subject>DNA - metabolism</subject><subject>DNA/dendrimer complexes</subject><subject>Electrophoresis, Agar Gel</subject><subject>Electrophoretic Mobility Shift Assay</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gel electrophoresis</subject><subject>Humans</subject><subject>Mice</subject><subject>Monophasic lysis</subject><subject>NIH 3T3 Cells</subject><subject>Polyamines - metabolism</subject><subject>Polypropylenes - metabolism</subject><subject>RNA - isolation & purification</subject><subject>RNA - metabolism</subject><subject>RNA isolation</subject><subject>Static Electricity</subject><subject>U937 Cells</subject><issn>0168-1656</issn><issn>1873-4863</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1P3DAQhq0KVJYtP6GVL_SWMI4dxzkhhApUQq20ggMny3EmrbeJvdjZSvz7mm4kjpxGeueZDz2EfGZQMmDyYltuOxdmtGUF0JTASuDwgayYanghlORHZJU5VTBZyxNymtIWAERbs4_khDVcSCXEijxtcPLGz9Sa2QXvLO3R99FNGBPtxmD_0M2PKzq5X_E_QJ2nOKKdY9j9DhGTS9QMM0Y6BZ8jk_KK8SXHn8jxYMaEZ0tdk8ebbw_Xd8X9z9vv11f3hRWVmAuBlRlqwKESHEHVXFnFBHYC-4aDUay3HFCKVjCulDID1kODvMvttrGV5Gvy9bB3F8PzHtOsJ5csjqPxGPZJN8AVyJq_C7K2AaZalcH6ANoYUoo46F0WYuKLZqBf5eutXuTrV_kamM7y89yX5cC-m7B_m1psZ-B8AUyyZhyi8dalN07JVkJdZe7ywGH29tdh1Mk69BZ7F7N63Qf3ziv_ACkMpgc</recordid><startdate>20070501</startdate><enddate>20070501</enddate><creator>Kuo, Jung-Hua Steven</creator><creator>Lin, Yi-Lin</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20070501</creationdate><title>Remnant cationic dendrimers block RNA migration in electrophoresis after monophasic lysis</title><author>Kuo, Jung-Hua Steven ; Lin, Yi-Lin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c424t-4e2af50ef243e08538c814eb4ed730a81dc30e649413888afe5f7e3bed797c263</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Cationic dendrimer</topic><topic>Dendrimers - metabolism</topic><topic>DNA - metabolism</topic><topic>DNA/dendrimer complexes</topic><topic>Electrophoresis, Agar Gel</topic><topic>Electrophoretic Mobility Shift Assay</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gel electrophoresis</topic><topic>Humans</topic><topic>Mice</topic><topic>Monophasic lysis</topic><topic>NIH 3T3 Cells</topic><topic>Polyamines - metabolism</topic><topic>Polypropylenes - metabolism</topic><topic>RNA - isolation & purification</topic><topic>RNA - metabolism</topic><topic>RNA isolation</topic><topic>Static Electricity</topic><topic>U937 Cells</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kuo, Jung-Hua Steven</creatorcontrib><creatorcontrib>Lin, Yi-Lin</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kuo, Jung-Hua Steven</au><au>Lin, Yi-Lin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Remnant cationic dendrimers block RNA migration in electrophoresis after monophasic lysis</atitle><jtitle>Journal of biotechnology</jtitle><addtitle>J Biotechnol</addtitle><date>2007-05-01</date><risdate>2007</risdate><volume>129</volume><issue>3</issue><spage>383</spage><epage>390</epage><pages>383-390</pages><issn>0168-1656</issn><eissn>1873-4863</eissn><coden>JBITD4</coden><abstract>Cationic dendrimers such as poly(amidoamine) (PAMAM) and poly(propyleneimine) (PPI) have attractive characteristics for the delivery of nucleic acid and various biomedical applications. Most studies have focused on cationic dendrimer-based intracellular delivery, and very few studies have focused on the non-specific interaction of remnant cationic dendrimers with total RNA after isolation directly from cells in vitro. We examined RNA isolation using the common method of monophasic lysis from human macrophage-like cells (U937) and mouse fibroblast cells (NIH/3T3) that had been exposed to dendrimers and DNA/dendrimer complexes using gel electrophoresis. We found that PAMAM and PPI dendrimers strongly altered the mobility of RNA in the gels. In addition, the extent of dendrimer-induced alteration in RNA mobility was directly dendrimer-generation-dependent: the alteration was greater with higher-generation dendrimers. We also found that DNA/dendrimer complexes at higher dendrimer to DNA ratios interacted with RNA after isolation while gene expression was maintained. The interactions between RNA and remnant dendrimers after isolation were caused by electrostatic bindings, and we recovered total RNA using high ionic strength solvents (2M NaCl solution) to disrupt the electrostatic forces binding dendrimers to RNA. Because RNA isolation is routinely used for biological applications, such dendrimer-induced alteration in RNA mobility should be accounted for in the further processing of RNA-related applications.</abstract><cop>Lausanne</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>17346844</pmid><doi>10.1016/j.jbiotec.2007.01.030</doi><tpages>8</tpages></addata></record> |
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subjects | Animals Biological and medical sciences Biotechnology Cationic dendrimer Dendrimers - metabolism DNA - metabolism DNA/dendrimer complexes Electrophoresis, Agar Gel Electrophoretic Mobility Shift Assay Fundamental and applied biological sciences. Psychology Gel electrophoresis Humans Mice Monophasic lysis NIH 3T3 Cells Polyamines - metabolism Polypropylenes - metabolism RNA - isolation & purification RNA - metabolism RNA isolation Static Electricity U937 Cells |
title | Remnant cationic dendrimers block RNA migration in electrophoresis after monophasic lysis |
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