Abstract
Antisense oligonucleotides (ASOs) are becoming important drugs for hard to treat diseases. Modifications to their DNA backbones are essential to inhibit degradation in vivo, but they can reduce binding affinity to RNA targets. To address this problem we have combined the enzymatic resistance of carbamate (CBM) DNA backbone analogues with the thermodynamic stability conferred by locked nucleic acid sugars (LNA). Using a dinucleotide phosphoramidite strategy and automated solid phase synthesis, we have synthesised a set of oligonucleotides modified with multiple LNA-CBM units. The LNA sugars restore binding affinity to RNA targets, and in this respect LNA position with respect to the CBM linkage is important. Oligonucleotides containing carbamate flanked on its 5'and 3'-sides by LNA form stable duplexes with RNA and unstable duplexes with DNA, which is desirable for antisense applications. Carbamate-LNA modified oligonucleotides also show increased stability in the presence of snake venom and foetal bovine serum compared to LNA or CBM backbones alone.
Highlights
Antisense oligonucleotides (ASOs) are short ∼20 mer sequences that bind to complementary ribonucleic nucleic acid (RNA) targets through Watson–Crick base pairing to promote enzymatic degradation (RNase H), induce RNA interference (RNAi) or regulate protein expression.[1]
Antisense oligonucleotides (ASOs) are becoming important drugs for hard to treat diseases. Modifications to their DNA backbones are essential to inhibit degradation in vivo, but they can reduce binding affinity to RNA targets. To address this problem we have combined the enzymatic resistance of carbamate (CBM) DNA backbone analogues with the thermodynamic stability conferred by locked nucleic acid sugars (LNA)
We report the synthesis of a set of dinucleotide phosphoramidites designed to introduce the carbamate-LNA modifications in Fig. 1 into oligonucleotides sequences using standard solid phase synthesis
Summary
Antisense oligonucleotides (ASOs) are short ∼20 mer sequences that bind to complementary ribonucleic nucleic acid (RNA) targets through Watson–Crick base pairing to promote enzymatic degradation (RNase H), induce RNA interference (RNAi) or regulate protein expression (splice modulation).[1]. To address this problem we have combined the enzymatic resistance of carbamate (CBM) DNA backbone analogues with the thermodynamic stability conferred by locked nucleic acid sugars (LNA). Carbamate-LNA modified oligonucleotides show increased stability in the presence of snake venom and foetal bovine serum compared to LNA or CBM backbones alone.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
