Synthesis and application of 2'-fluoro-substituted cap analogs.
Design, synthesis, and biological evaluation of 2'-fluoro-substituted cap analogs, i.e., m(7,2'F)G[5']ppp[5']G and m(7,2'F)G[5']ppp[5']m(7)G are described. Structures were confirmed by (1)H, (31)P, (19)F NMR and MS data. The effects of the 2'-fluoro-substituted moiety from the normal and N(7) double methylated mCAP were evaluated with respect to their capping efficiency, in vitro T7 RNA polymerase transcription efficiency, and translation activity using cultured HeLa cells. Luciferase fusion protein production was monitored by measuring the luciferase activity. mRNA poly(A) capped with 2'-fluoro-substituted cap analogs, (m(7,2'F)G[5']ppp[5']G) and (m(7,2'F)G[5']ppp[5']m(7)G), were translated approximately 2.4- and 2.5-fold more efficiently, respectively, than mRNA capped with conventional m(7)G[5']ppp[5']G.
Kore
Anilkumar R
AR
Ambion Inc., An Applied Biosystems Business, Bioorganic Chemistry Division, 2130 Woodward Street, Austin, TX 78744-1832, USA. anil.kore@appliedbiosystems.com
Shanmugasundaram
Muthian
M
Charles
Irudaya
I
Cheng
Angie M
AM
Barta
Timothy J
TJ
D000199:Actins / Q000096:biosynthesis / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors*; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000138:chemical synthesis* / Q000494:pharmacology*; D017930:Genes, Reporter / Q000235:genetics; D006151:Guanosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D013329:Structure-Activity Relationship; D014764:Viral Proteins / Q000037:antagonists & inhibitors*
C116616:8-methylguanosine; D000199:Actins; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D014764:Viral Proteins; D006151:Guanosine; D008156:Luciferases; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases