ABSTRACT
Isatoic anhydride derivatives, including a biotin and a disulfide linker were specifically designed for nucleic acid separation. 2'-OH selective RNA acylation, capture of biotinylated RNA adducts by streptavidin-coated magnetic beads and disulfide chemical cleavage led to isolation of highly enriched RNA samples from an initial 9/1 DNA-RNA mixture. Starting from the parent compound N-methylisatoic anhydride A which was used at 65 °C, we improved the extraction process by designing a new generation of isatoic anhydrides that are able to react under smoother conditions. Among them, a pyridine-based isatoic anhydride derivative 15f was found to be reactive at room temperature, leading to enhance the efficiency and selectivity of the extraction process by significantly reducing DNA side extraction. The extracted and purified RNAs can then be detected by RT-PCR.
Subject(s)
Biotin/chemistry , Oxazines/chemistry , Pyridines/chemistry , RNA/isolation & purification , Temperature , Acylation , Chromatography, Liquid , DNA/chemistry , Esters/chemical synthesis , Esters/chemistry , HIV/genetics , Mass Spectrometry , Oxazines/chemical synthesis , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Viral/genetics , RNA, Viral/isolation & purification , Real-Time Polymerase Chain ReactionABSTRACT
An isatoic anhydride derivative conjugated to a biotin and a disulfide linker was specifically designed for the separation of nucleic acids. Starting from a DNA-RNA mixture, a selective 2'-hydroxyl acylation of RNAs followed by capture with streptavidin-coated magnetic beads and cleavage of the disulfide led to elution of RNAs.
Subject(s)
Anhydrides/chemistry , Biotin/chemistry , DNA/chemistry , DNA/isolation & purification , Hydroxides/chemistry , RNA/chemistry , RNA/isolation & purification , ortho-Aminobenzoates/chemistry , Acylation , RNA, Viral/chemistry , RNA, Viral/isolation & purification , Streptavidin/chemistryABSTRACT
Using solid phase synthesis techniques, we have rapidly obtained a series of eight aryl sulphonamides derived from putrescine. These conjugates with various aryl groups were evaluated for their affinity towards 5-HT6 receptors in man. This evaluation revealed the interest of two compounds which present the same activity level, in the submicromolar range, as two reference derivatives. The most potent will be considered as a new lead for further investigations.
