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10.1021/jacs.1c00014 http://scihub22266oqcxt.onion/10.1021/jacs.1c00014 33797236!8154517!33797236     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=33797236&cmd=llinks): Failed to open stream: HTTP request failed! HTTP/1.1 429 Too Many Requests in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 215 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       J+Am+Chem+Soc 2021 ; 143 (14): 5413-5424 Nephropedia Template TP {{tp|p=33797236|t=2021. Stealth Fluorescence Labeling for Live Microscopy Imaging of mRNA Delivery |pdf=|usr=}}{{33797236}} gab.com Text Stealth Fluorescence Labeling for Live Microscopy Imaging of mRNA Delivery JO: J Am Chem Soc http://www.kidney.de/pget.php?&tw=1&pmid=33797236 #FOAvip Methods for tracking RNA inside living cells without perturbing their natural interactions and functions are critical within biology and, in particular, to facilitate studies of therapeutic RNA delivery. We present a stealth labeling approach that can efficiently, and with high fidelity, generate RNA transcripts, through enzymatic incorporation of the triphosphate of tC(O), a fluorescent tricyclic cytosine analogue. We demonstrate this by incorporation of tC(O) in up to 100% of the natural cytosine positions of a 1.2 kb mRNA encoding for the histone H2B fused to GFP (H2B:GFP). Spectroscopic characterization of this mRNA shows that the incorporation rate of tC(O) is similar to cytosine, which allows for efficient labeling and controlled tuning of labeling ratios for different applications. Using live cell confocal microscopy and flow cytometry, we show that the tC(O)-labeled mRNA is efficiently translated into H2B:GFP inside human cells. Hence, we not only develop the use of fluorescent base analogue labeling of nucleic acids in live-cell microscopy but also, importantly, show that the resulting transcript is translated into the correct protein. Moreover, the spectral properties of our transcripts and their translation product allow for their straightforward, simultaneous visualization in live cells. Finally, we find that chemically transfected tC(O)-labeled RNA, unlike a state-of-the-art fluorescently labeled RNA, gives rise to expression of a similar amount of protein as its natural counterpart, hence representing a methodology for studying natural, unperturbed processing of mRNA used in RNA therapeutics and in vaccines, like the ones developed against SARS-CoV-2. Twit Text FOAVip Stealth Fluorescence Labeling for Live Microscopy Imaging of mRNA Delivery ????J Am Chem Soc http://www.kidney.de/pget.php?&tw=1&pmid=33797236 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33797236 #FOAvip English Wikipedia <ref>{{Cite pmid|33797236}}</ref> Stealth Fluorescence Labeling for Live Microscopy Imaging of mRNA Delivery #MMPMID33797236 Baladi T; Nilsson JR; Gallud A; Celauro E; Gasse C; Levi-Acobas F; Sarac I; Hollenstein MR; Dahlen A; Esbjorner EK; Wilhelmsson LM J Am Chem Soc 2021[Apr]; 143 (14): 5413-5424 PMID33797236show ga Methods for tracking RNA inside living cells without perturbing their natural interactions and functions are critical within biology and, in particular, to facilitate studies of therapeutic RNA delivery. We present a stealth labeling approach that can efficiently, and with high fidelity, generate RNA transcripts, through enzymatic incorporation of the triphosphate of tC(O), a fluorescent tricyclic cytosine analogue. We demonstrate this by incorporation of tC(O) in up to 100% of the natural cytosine positions of a 1.2 kb mRNA encoding for the histone H2B fused to GFP (H2B:GFP). Spectroscopic characterization of this mRNA shows that the incorporation rate of tC(O) is similar to cytosine, which allows for efficient labeling and controlled tuning of labeling ratios for different applications. Using live cell confocal microscopy and flow cytometry, we show that the tC(O)-labeled mRNA is efficiently translated into H2B:GFP inside human cells. Hence, we not only develop the use of fluorescent base analogue labeling of nucleic acids in live-cell microscopy but also, importantly, show that the resulting transcript is translated into the correct protein. Moreover, the spectral properties of our transcripts and their translation product allow for their straightforward, simultaneous visualization in live cells. Finally, we find that chemically transfected tC(O)-labeled RNA, unlike a state-of-the-art fluorescently labeled RNA, gives rise to expression of a similar amount of protein as its natural counterpart, hence representing a methodology for studying natural, unperturbed processing of mRNA used in RNA therapeutics and in vaccines, like the ones developed against SARS-CoV-2. |*Fluorescence[MESH] |*Molecular Imaging[MESH] |COVID-19 Drug Treatment[MESH] |Cell Line, Tumor[MESH] |Cytosine/analogs & derivatives/analysis/chemical synthesis/chemistry[MESH] |Fluorescent Dyes/*analysis/chemical synthesis/*chemistry[MESH] |Green Fluorescent Proteins/metabolism[MESH] |Histones/metabolism[MESH] |Humans[MESH] |Molecular Structure[MESH] |RNA, Messenger/*analysis/chemistry/*metabolism/therapeutic use[MESH]Stealth Fluorescence Labeling for Live Microscopy Imaging of mRNA Deli(epmc).pdf DeepDyve Pubget Overpricing