Synthesis and evaluation of modified siRNA molecules containing a novel glucose derivative

Chemical modifications are critical for the development of safe and effective siRNAs for downstream applications. In this study, we report the synthesis of a novel glucose phosphoramidite, a triazole-linked to uracil at position one, for incorporation into oligonucleotides. Biological testing revealed that the glucose derivative at key positions within the sense or antisense strand can lead to potent gene-silencing activity, thus highlighting its tolerance in both sense and antisense positions. Furthermore, the A-form helical formation was maintained with this modification. Overall, placing the modification at the 3′ end and at key internal positions led to effective RNAi gene-silencing activity.

[1]  M. Manoharan,et al.  Chimeric siRNAs with chemically modified pentofuranose and hexopyranose nucleotides: altritol-nucleotide (ANA) containing GalNAc–siRNA conjugates: in vitro and in vivo RNAi activity and resistance to 5′-exonuclease , 2020, Nucleic acids research.

[2]  L. Scott.  Givosiran: First Approval , 2020, Drugs.

[3]  S. Solomon,et al.  Patisiran, an RNAi Therapeutic, for Hereditary Transthyretin Amyloidosis , 2018, The New England journal of medicine.

[4]  K. Ranganna,et al.  Therapeutic potential of chemically modified siRNA: Recent trends , 2017, Chemical biology & drug design.

[5]  D. Haussecker.  Current issues of RNAi therapeutics delivery and development. , 2014, Journal of controlled release : official journal of the Controlled Release Society.

[6]  Jian Zhou,et al.  Regiospecific anomerisation of acylated glycosyl azides and benzoylated disaccharides by using TiCl4. , 2013, Chemistry.

[7]  K. Ye,et al.  Single modification at position 14 of siRNA strand abolishes its gene‐silencing activity by decreasing both RISC loading and target degradation , 2013, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[8]  Jesper B. Bramsen,et al.  Development of Therapeutic-Grade Small Interfering RNAs by Chemical Engineering , 2012, Front. Gene..

[9]  E. Kool,et al.  Steric restrictions of RISC in RNA interference identified with size-expanded RNA nucleobases. , 2012, ACS chemical biology.

[10]  A. Willingham,et al.  siRNA-optimized Modifications for Enhanced In Vivo Activity , 2012, Molecular therapy. Nucleic acids.

[11]  Daniel Shriner,et al.  Moving toward System Genetics through Multiple Trait Analysis in Genome-Wide Association Studies , 2011, Front. Gene..

[12]  P. Herdewijn,et al.  Biological effects of hexitol and altritol-modified siRNAs targeting B-Raf. , 2009, European journal of pharmacology.

[13]  Jaroslav Kypr,et al.  Circular dichroism and conformational polymorphism of DNA , 2009, Nucleic acids research.

[14]  J. Watts,et al.  Chemically modified siRNA: tools and applications. , 2008, Drug discovery today.

[15]  P. Herdewijn,et al.  Structural characterization and biological evaluation of small interfering RNAs containing cyclohexenyl nucleosides. , 2007, Journal of the American Chemical Society.

[16]  J. Rossi,et al.  RNAi therapeutics: principles, prospects and challenges. , 2007, Advanced drug delivery reviews.

[17]  P. Herdewijn,et al.  Inhibition of MDR1 expression with altritol-modified siRNAs , 2007, Nucleic acids research.

[18]  M. Serra,et al.  Stability of 3' double nucleotide overhangs that model the 3' ends of siRNA. , 2005, RNA.

[19]  C. Wahlestedt,et al.  Locked nucleic acid (LNA) mediated improvements in siRNA stability and functionality , 2005, Nucleic acids research.

[20]  R. Bhatnagar,et al.  RNA Interference: Biology, Mechanism, and Applications , 2003, Microbiology and Molecular Biology Reviews.

[21]  T. Rana,et al.  siRNA function in RNAi: a chemical modification analysis. , 2003, RNA.

[22]  David R Corey,et al.  RNA interference in mammalian cells by chemically-modified RNA. , 2003, Biochemistry.

[23]  G. Hannon.  RNA interference : RNA , 2002 .

[24]  Gregory J. Hannon,et al.  Insight Review Articles , 2022 .

[25]  T. Tuschl,et al.  Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells , 2001, Nature.

[26]  J. Wengel,et al.  Synthesis and evaluation of oligodeoxynucleotides containing acyclic nucleosides: introduction of three novel analogues and a summary. , 1995, Bioorganic & medicinal chemistry.