A5051368682- RNA and protein synthesis mechanisms
- RNA Research and Splicing
- RNA modifications and cancer
- RNA Interference and Gene Delivery
- Click Chemistry and Applications
- Cancer-related gene regulation
- Advanced biosensing and bioanalysis techniques
- DNA and Nucleic Acid Chemistry
- RNA regulation and disease
- Viral Infections and Immunology Research
- Virus-based gene therapy research
- Bacteriophages and microbial interactions
- Trypanosoma species research and implications
- Research on Leishmaniasis Studies
- Poxvirus research and outbreaks
- interferon and immune responses
- Chemical Synthesis and Analysis
- Carbohydrate Chemistry and Synthesis
- History and advancements in chemistry
- Freezing and Crystallization Processes
- Various Chemistry Research Topics
- Fluorine in Organic Chemistry
- Biochemical and Molecular Research
- Synthesis and Reactions of Organic Compounds
- Advanced NMR Techniques and Applications
University of Warsaw
2016-2025
Institute of Experimental Physics of the Slovak Academy of Sciences
2016-2024
Abstract 7-Methylguanosine 5′ cap on mRNA is necessary for efficient protein expression in vitro and vivo. Recent studies revealed structural diversity of endogenous caps, which carry different 5′-terminal nucleotides additional methylations (2′-O-methylation m6A). Currently available 5′-capping methods do not address this diversity. We report trinucleotide analogs (m7GpppN(m)pG), are utilized by RNA polymerase T7 to initiate transcription from templates carrying Φ6.5 promoter enable...
Abstract In mammals, m7G-adjacent nucleotides undergo extensive modifications. Ribose of the first or and second transcribed can be subjected to 2′-O-methylation form cap1 cap2, respectively. When nucleotide is 2′-O-methylated adenosine, it additionally modified N6,2′-O-dimethyladenosine (m6Am). Recently, crucial role in distinguishing between ‘self’ ‘non-self’ mammalian cells during viral infection was revealed. Here, we attempted understand impact cap methylations on RNA-related processes....
ConspectusMessenger ribonucleic acid (mRNA) is the universal cellular instruction for ribosomes to produce proteins. Proteins are responsible most of functions living organisms, and their abnormal structure or activity cause many diseases. mRNA, which expressed in cytoplasm and, unlike DNA, does not need be delivered into nucleus, appears an ideal vehicle pursuing idea gene therapy genetic information about proteins introduced organism exert a therapeutic effect. mRNA molecules any sequence...
Eukaryotic mRNAs undergo cotranscriptional 5′-end modification with a 7-methylguanosine cap. In higher eukaryotes, the cap carries additional methylations, such as m6Am─a common epitranscriptomic mark unique to mRNA 5′-end. This is regulated by Pcif1 methyltransferase and FTO demethylase, but its biological function still unknown. Here, we designed synthesized trinucleotide FTO-resistant N6-benzyl analogue of m6Am-cap–m7GpppBn6AmpG (termed AvantCap) incorporated it into using T7 polymerase....
The 5′ cap consists of 7-methylguanosine (m7G) linked by a 5′–5′-triphosphate bridge to messenger RNA (mRNA) and acts as the master regulator mRNA turnover translation initiation in eukaryotes. Cap analogues that influence (either small molecules or part an transcript) are valuable tools for studying gene expression, which is often also therapeutic relevance. Here, we synthesized series 15 dinucleotide (m7GpppG) containing 5′-phosphorothiolate (5′-PSL) moiety (i.e., O-to-S substitution...
The m7G cap is ubiquitous on RNAPII-transcribed RNA and has fundamental roles in eukaryotic gene expression, however its vivo role mammals remained unknown. Here, we identified the methyltransferase, RNMT, as a key mediator of T cell activation, which specifically regulates ribosome production. During induction mRNA expression biogenesis drives metabolic reprogramming, rapid proliferation differentiation generating effector populations. We report that RNMT induced by receptor (TCR)...
Chemical modifications enable preparation of mRNAs with augmented stability and translational activity. In this study, we explored how chemical 5′,3′-phosphodiester bonds in the mRNA body poly(A) tail influence biological properties eukaryotic mRNA. To obtain modified unmodified vitro transcribed mRNAs, used ATP analogs at α-phosphate (containing either O-to-S or O-to-BH 3 substitutions) three different RNA polymerases—SP6, T7, polymerase. verify efficiency incorporation presence ATP,...
Chemical modification of messenger RNA (mRNA) has paved the way for advancing mRNA-based therapeutics. The intricate process mRNA translation in eukaryotes is orchestrated by numerous proteins involved complex interaction networks. Many them bind specifically to a unique structure at 5'-end, called 5'-cap. Depending on 5'-terminal sequence and its methylation pattern, different may be initiation regulation, but deeper understanding these mechanisms requires specialized molecular tools...
mRNA-based gene delivery is a powerful strategy for many therapeutic areas. In this work, we used CuAAC to synthesize next-generation triazole-bearing mRNA 5' cap analogs and evaluated them as reagents modification of in vitro transcribed mRNA.
To broaden the scope of existing methods based on (19)F nucleotide labeling, we developed a new method for synthesis fluorophosphate (oligo)nucleotide analogues containing an O to F substitution at terminal position (oligo)phosphate moiety and evaluated them as tools NMR studies. Using three efficient comprehensive synthetic approaches phosphorimidazolide chemistry tetra-n-butylammonium fluoride, fluoromonophosphate, or imidazolide fluorine sources, prepared over 30...
Analogues of the mRNA 5'-cap are useful tools for studying translation and degradation, with emerging potential applications in novel therapeutic interventions including gene therapy. We report synthesis mono- dinucleotide cap analogues containing dihalogenmethylenebisphosphonate moiety (i.e. one bridging O atom substituted CCl2 or CF2) their properties context cellular translational decapping machineries, compared to phosphate-unmodified previously reported CH2-substituted caps. The were...
Abstract The 7‐methylguanosine (m 7 G) cap structure is a unique feature present at the 5′ ends of messenger RNAs (mRNAs), and it can be subjected to extensive modifications, resulting in alterations mRNA properties (e.g. translatability, susceptibility degradation). It also provide molecular tools study metabolism. We developed new analogues that enable site‐specific labeling RNA end using strain‐promoted azide–alkyne cycloaddition (SPAAC) without disrupting basic function protein...
mRNA-based therapies and vaccines constitute a disruptive technology with the potential to revolutionize modern medicine. Chemically modified 5' cap structures have provided access mRNAs superior translational properties that could benefit currently flourishing mRNA field. Prime examples of compounds enhance are antireverse analog diastereomers contain an O-to-S substitution within β-phosphate (β-S-ARCA D1 D2), where is used in clinically investigated vaccines. The were previously found high...
The high sensitivity of 19F nucleus to changes in the chemical environment has promoted use fluorine-labeled molecular probes study structure and interactions nucleic acids by NMR. So far, most efforts have focused on incorporating fluorine atom into nucleobase ribose moieties using either monomer building blocks for solid-phase synthesis, or nucleoside triphosphates enzymatic synthesis. Here, we report a simple efficient synthesis 5'-fluoromonophosphorylated 5'-fluorodiphosphorylated...
mRNA is a template for protein biosynthesis, and consequently transport, translation, turnover are key elements in the overall regulation of gene expression. Along with growing interest mechanisms regulating decay localization, there an increasing need tools enabling convenient fluorescent labeling or affinity tagging mRNA. We report new 5' cap analog-based that enable site-specific RNA within using N-hydroxysuccinimide (NHS) chemistry. explored two complementary methods: co-transcriptional...
Herein, we report a straightforward one-step procedure for modifying N-nucleophilic groups in the nucleobases of commercially available nucleoside phosphoramidites. This method involves deprotonation amide under phase-transfer conditions and subsequent reaction with electrophilic molecules such as alkyl halides or organic isocyanates. Using this approach, obtained 10 different classes modified phosphoramidites suitable synthesis oligonucleotides, including several noncanonical nucleotides...
We report the chemical synthesis of a set nicotinamide adenine dinucleotide (NAD) cap analogues containing modifications that reduce their susceptibility to NAD-RNA-degrading enzymes. These can be incorporated into transcripts in similar way as NAD. Biochemical characterization RNAs carrying these caps with DXO, NudC, and Nudt12 enzymes led identification compounds instrumental unraveling so far unaddressed biological aspects NAD-RNAs.
In response to foreign RNA, cellular antiviral mechanisms stimulate high expression of interferon-induced proteins with tetratricopeptide repeats (IFITs). Two members the IFIT protein family, IFIT1 and IFIT5, are capable binding very terminal 5′ end mRNA. eukaryotes, these mRNA termini contain a cap structure (m 7 GpppN, 0) that is often subjected further modifications. Here, we performed thorough examination IFIT5 wide spectrum differently capped as well fully uncapped mRNAs. The kinetic...
Abstract Kinetoplastids are a clade of eukaryotic protozoans that include human parasitic pathogens like trypanosomes and Leishmania species. In these organisms, protein-coding genes transcribed as polycistronic pre-mRNAs, which need to be processed by the coupled action trans-splicing polyadenylation yield monogenic mature mRNAs. During trans-splicing, universal RNA sequence, spliced leader (SL RNA) mini-exon, is added 5’-end each mRNA. The this mini-exon carries hypermethylated cap...
Commercially available 2′-O-pivaloyloxymethyl (PivOM) phosphoramidites were employed in an SPS protocol for RNA 5′ azides. The utility of the N3-RNAs CuAAC and SPAAC was demonstrated by labeling, chemical ligation including fragment joining cyclization, bioconjugation. As a result, several new conjugates that may be valuable tools studies on biological events such as innate immune response (cyclic dinucleotides), post-transcriptional gene regulation (circular RNAs), or mRNA turnover (m7G...
Phosphate-modified m<sub>3</sub>G cap analogs were synthesized, conjugated to RNA using “click chemistry”, and studied for susceptibility hNUDT16 enzyme.