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- W4294591381 endingPage "e115" @default.
- W4294591381 startingPage "e115" @default.
- W4294591381 abstract "Abstract Synthetic mRNA has recently moved into the focus of therapeutic and vaccination efforts. Incorporation of modified nucleotides during in vitro transcription can improve translation and attenuate immunogenicity, but is limited to triphosphate nucleotides which are accepted by RNA polymerases, and their incorporation is either random or complete. In contrast, site-specific modification, herein termed ‘point modification’ in analogy to point mutations, holds significant technical challenge. We developed fundamental techniques for isolation of long, translatable and internally point-modified mRNAs. Enabling concepts include three-way-one-pot splint ligations, and isolation of mRNA by real-time elution from agarose gels. The use of blue light permitted visualization of mRNA in pre-stained gels without the photochemical damage associated with the use of hard UV-radiation. This allowed visualization of the mRNA through its migration in the agarose gel, which in turn, was a prerequisite for its recovery by electroelution into precast troughs. Co-eluting agarose particles were quantified and found to not be detrimental to mRNA translation in vitro. Translation of EGFP-coding mRNA into functional protein was quantified by incorporation of 35S-labelled methionine and by in-gel EGFP fluorescence. This enabled the functional analysis of point modifications, specifically of ribose methylations in the middle of a 1371 nt long mRNA." @default.
- W4294591381 created "2022-09-05" @default.
- W4294591381 creator A5001118364 @default.
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- W4294591381 creator A5089150142 @default.
- W4294591381 date "2022-09-05" @default.
- W4294591381 modified "2023-10-17" @default.
- W4294591381 title "Synthesis of point-modified mRNA" @default.
- W4294591381 cites W1215958058 @default.
- W4294591381 cites W127850330 @default.
- W4294591381 cites W1484258617 @default.
- W4294591381 cites W1498804507 @default.
- W4294591381 cites W1538766821 @default.
- W4294591381 cites W1569097650 @default.
- W4294591381 cites W1579335652 @default.
- W4294591381 cites W1591867940 @default.
- W4294591381 cites W1788436564 @default.
- W4294591381 cites W1923876136 @default.
- W4294591381 cites W1963706689 @default.
- W4294591381 cites W1967267862 @default.
- W4294591381 cites W1969918693 @default.
- W4294591381 cites W1986178107 @default.
- W4294591381 cites W1991807334 @default.
- W4294591381 cites W1994391392 @default.
- W4294591381 cites W1997906712 @default.
- W4294591381 cites W1998059339 @default.
- W4294591381 cites W2002376867 @default.
- W4294591381 cites W2016786790 @default.
- W4294591381 cites W2017505216 @default.
- W4294591381 cites W2017515700 @default.
- W4294591381 cites W2018134130 @default.
- W4294591381 cites W2019990686 @default.
- W4294591381 cites W2023169739 @default.
- W4294591381 cites W2024227631 @default.
- W4294591381 cites W2025980739 @default.
- W4294591381 cites W2029691070 @default.
- W4294591381 cites W2030673136 @default.
- W4294591381 cites W2031061885 @default.
- W4294591381 cites W2032579724 @default.
- W4294591381 cites W2039243132 @default.
- W4294591381 cites W2039422212 @default.
- W4294591381 cites W2040651187 @default.
- W4294591381 cites W2041914510 @default.
- W4294591381 cites W2050843600 @default.
- W4294591381 cites W2054248438 @default.
- W4294591381 cites W2058647478 @default.
- W4294591381 cites W2077891946 @default.
- W4294591381 cites W2082431303 @default.
- W4294591381 cites W2083805819 @default.
- W4294591381 cites W2083958143 @default.
- W4294591381 cites W2085749025 @default.
- W4294591381 cites W2088873069 @default.
- W4294591381 cites W2091039881 @default.
- W4294591381 cites W2098597036 @default.
- W4294591381 cites W2100211480 @default.
- W4294591381 cites W2106311383 @default.
- W4294591381 cites W2108332331 @default.
- W4294591381 cites W2115728811 @default.
- W4294591381 cites W2117229732 @default.
- W4294591381 cites W2118598894 @default.
- W4294591381 cites W2123644491 @default.
- W4294591381 cites W2123848310 @default.
- W4294591381 cites W2127068865 @default.
- W4294591381 cites W2137186478 @default.
- W4294591381 cites W2137450588 @default.
- W4294591381 cites W2142739728 @default.
- W4294591381 cites W2147363872 @default.
- W4294591381 cites W2162970812 @default.
- W4294591381 cites W2168071482 @default.
- W4294591381 cites W2169838876 @default.
- W4294591381 cites W2175514210 @default.
- W4294591381 cites W2190510612 @default.
- W4294591381 cites W2246990966 @default.
- W4294591381 cites W2340049980 @default.
- W4294591381 cites W2412158473 @default.
- W4294591381 cites W2413444844 @default.
- W4294591381 cites W2418104675 @default.
- W4294591381 cites W2419188398 @default.
- W4294591381 cites W2432150382 @default.
- W4294591381 cites W2474169621 @default.
- W4294591381 cites W2492904803 @default.
- W4294591381 cites W2551836638 @default.
- W4294591381 cites W2584249987 @default.
- W4294591381 cites W2587850379 @default.
- W4294591381 cites W2589945527 @default.
- W4294591381 cites W2590034108 @default.
- W4294591381 cites W2593990758 @default.
- W4294591381 cites W2606518593 @default.
- W4294591381 cites W2612024267 @default.
- W4294591381 cites W2621631583 @default.
- W4294591381 cites W2744180694 @default.
- W4294591381 cites W2768377421 @default.
- W4294591381 cites W2781829910 @default.
- W4294591381 cites W2783301206 @default.