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10.1093/nar/gkv307 http://scihub22266oqcxt.onion/10.1093/nar/gkv307 C4482072!4482072 !25870407     free     free     free Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\25870407 .jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117       Nucleic+Acids+Res 2015 ; 43 (9 ): 4650-60 Nephropedia Template TP {{tp|p=25870407 |t=2015. Live cell imaging of duplex siRNA intracellular trafficking |pdf=|usr=}}{{25870407 }} gab.com Text Live cell imaging of duplex siRNA intracellular trafficking JO: Nucleic Acids Res http://www.kidney.de/pget.php?&tw=1&pmid=25870407 #FOAvip Intracellular distribution of siRNA after in vitro transfection typically depends on lipopolyplexes, which must release the siRNA into the cytosol. Here, the fate of siRNAs was monitored by FRET-based live cell imaging. Subsequent to in situ observation of uptake and release processes, this approach allowed the observation of a number of hitherto uncharacterized intracellular distribution and degradation processes, commencing with a burst of endosomal releases, followed, in some cases, by fast siRNA influx into the nucleus. The continued observation of intact siRNA against a background of free fluorophores resulting from advanced degradation was possible by a specifically developed imaging algorithm, which identified populations of intact siRNA in pixels based on FRET. This proved to be essential in the end point definition of siRNA distribution, which typically featured partially degraded siRNA pools in perinuclear structures. Our results depict the initial 4 h as a critical time window, characterized by fast initial burst release into the cytosol, which lay the foundations for subsequent intracellular distribution of siRNA. Combination with a subsequent slower, but sustained release from endosomal reservoirs may contribute to the efficiency and duration of RNAi, and explain the success of lipopolyplexes in RNAi experiments in cell culture. Twit Text FOAVip Live cell imaging of duplex siRNA intracellular trafficking ????Nucleic Acids Res http://www.kidney.de/pget.php?&tw=1&pmid=25870407 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=25870407 #FOAvip English Wikipedia <ref>{{Cite pmid|25870407 }}</ref> Live cell imaging of duplex siRNA intracellular trafficking #MMPMID25870407 Hirsch M ; Helm M Nucleic Acids Res 2015[May]; 43 (9 ): 4650-60 PMID25870407 show ga Intracellular distribution of siRNA after in vitro transfection typically depends on lipopolyplexes, which must release the siRNA into the cytosol. Here, the fate of siRNAs was monitored by FRET-based live cell imaging. Subsequent to in situ observation of uptake and release processes, this approach allowed the observation of a number of hitherto uncharacterized intracellular distribution and degradation processes, commencing with a burst of endosomal releases, followed, in some cases, by fast siRNA influx into the nucleus. The continued observation of intact siRNA against a background of free fluorophores resulting from advanced degradation was possible by a specifically developed imaging algorithm, which identified populations of intact siRNA in pixels based on FRET. This proved to be essential in the end point definition of siRNA distribution, which typically featured partially degraded siRNA pools in perinuclear structures. Our results depict the initial 4 h as a critical time window, characterized by fast initial burst release into the cytosol, which lay the foundations for subsequent intracellular distribution of siRNA. Combination with a subsequent slower, but sustained release from endosomal reservoirs may contribute to the efficiency and duration of RNAi, and explain the success of lipopolyplexes in RNAi experiments in cell culture. |Animals [MESH] |Cell Line [MESH] |Cell Nucleus/metabolism [MESH] |Endosomes/metabolism [MESH] |Fluorescence Resonance Energy Transfer [MESH] |Microscopy, Confocal [MESH] |RNA Transport [MESH] |RNA, Small Interfering/*metabolism [MESH] |Rats [MESH]Live cell imaging of duplex siRNA intracellular trafficking (epmc).pdf DeepDyve Pubget Overpricing