Designing mRNA transcripts resistant to enzymatic cap hydrolysis applicable for anti-cancer vaccines; multiplexed biochemical and biophysical studies on representative classes of decapping enzymes.
2013/08/A/NZ1/00866
Keywords
- 5' mRNA monomethylguanosine cap
- trimethylguanosine cap
- cap-4
- decapping enzymes
- HIT hydrolases
- NUDIX hydrolases
- human DcpS
- yeast Dcs1
- C.elegans DcpS
- Nudt16
- yeast Dcp2/Dcp1
- Trypanosomatide HIT-45
- translation initiation factor eIF4E
- fluorescence titration TST
- Reverse Phase HPLC
- isothermal calorimetric titration ITC
- near UV circular dichroism CD
- stopped flow
- kinetic and thermodynamic parameters
- molecular modeling
- NMR spectroscopy
- mass spectroscopy
- non-hydrolyzable synthetic cap analogues
- anti-tumor mRNA vaccinies
- anti reverse cap analogues ARCAs
- phosphate bridge modifications
- alternatively capped oligonucleotides and mRNAs
- chemoenzymatic mRNA capping
- protein translation
- translation inhibition
Descriptors
- NZ1_1: Biologia molekularna
- NZ1_3: Biofizyka
- NZ1_6: Biologia syntetyczna
Panel
NZ1 – Molecular biology, structural biology, biotechnology: molecular biology, structural biology, biotechnology
Host institution
Uniwersytet Warszawski
Voivodeship: mazowieckie
Principal investigator (from the host institution)
Number of co-investigators in the project: 20
Call
MAESTRO 4 – announced on 2012-12-15
- Amount awarded
- 2 983 800 PLN
- Project start date
- 2013-09-23
- Project end date
- 2019-09-22
- Project duration
- 72 months (the same as in the proposal)
- Project status
- Project settled
Equipment purchased [PL]
- Aparat HPLC. Cost: 190 000 PLN
- Aparat do badania oddziaływań pomiędzy makrocząsteczkami.
- Chłodziarko-zamrażarki.
- Kolektor frakcji. Cost: 32 000 PLN
- Komputery.
- Komputery, oprogramowanie biurowe i specjalistyczne oraz akcesoria komputerowe. Cost: 7 226 PLN
- Koszyki do wirowania płytek. Cost: 7 131 PLN
- Luminometr. Cost: 38 000 PLN
- Pipeta 8-kanałowa o zmiennych pojemnościach z pudełkami na końcówki. Cost: 2 484 PLN
- Podręczny mikrofalowy autoklaw.
- Spektrofluorymetr. Cost: 220 000 PLN
- Spektrofotometr dwuwiązkowy UV-VIS.
- Witryna chłodnicza.
- Wymienny termoblok na probówki stożkowe. Cost: 2 329 PLN
- Wyparka próżniowa.
- Wózki do pożywek.
Information in the final report
Publication in academic press/journals (19)
-
Structural analysis of human 2'-O-ribose methyltransferases involved in mRNA cap structure formation
- Authors:
- Smietanski M., Werner M., Purta E., Kaminska K.H., Stepinski J., Darzynkiewicz E., Nowotny M., Bujnicki J.M.
- Academic press:
- Nature Communications (year: 2014, volume: 5, pages: 46032), Publisher: Macmillan Publishers Limited
- Status:
- Published
- DOI:
- 10.1038/ncomms4004 – link to the publication
-
Synthesis, properties, and biological activity of boranophosphate analogs of the mRNA cap: versalite tools for manipulation of therapeutically relevant cap-dependent processes
- Authors:
- Kowalska J., Wypijewska del Nogal A., Darzynkiewicz Z.M., Buck J., Nicola C., Kuhn A.N., Lukaszewicz M., Zuberek J., Strenkowska M., Ziemniak M., Maciejczyk M., Bojarska E., Rhoads R.E., Darzynkiewicz E., Sahin U., Jemielity J.
- Academic press:
- Nucleic Acids Research (year: 2014, volume: 42, pages: 10245-10264), Publisher: Oxford University Press
- Status:
- Published
- DOI:
- 10.1093/nar/gku757 – link to the publication
-
Triazole-containing monophosphate mRNA cap analogs as effective translation inhibitors
- Authors:
- Piecyk K., Lukaszewicz M., Darzynkiewicz E., Jankowska-Anyszka M.
- Academic press:
- RNA (year: 2014, volume: 20, pages: 1539-1547), Publisher: Cold Spring Harbor Laboratory Press
- Status:
- Published
- DOI:
- 10.1261/rna.046193.114 – link to the publication
-
Effect of the His-Tag Location on Decapping Scavenger Enzymes and Their Hydrolytic Activity toward Cap Analogs
- Authors:
- Ferenc-Mrozek A., Bojarska E., Stepinski J., Darzynkiewicz E., Lukasiewicz M.
- Academic press:
- ACS Omega (year: 2020, volume: 5, pages: 10759-10766), Publisher: ACS Publications
- Status:
- Published
- DOI:
- 10.1021/acsomega.0c00304 – link to the publication
-
Kinetic analysis of IFIT1 and IFIT5 interaction with different native and engineered RNAs and its consequences for designing mRNA-based therapeutics.
- Authors:
- Miedziak B., Dobiezynska A., Darzynkiewicz Z.M., Bartkowska J., Miszkiewicz J., Kowalska J., Warminski M., Tyras M., Trylska J., Jemielity J., Darzynkiewicz E., Grzela R.
- Academic press:
- RNA (year: 2020, volume: 26, pages: 58-68), Publisher: Cold Spring Harbor Laboratory Press
- Status:
- Published
- DOI:
- 10.1261/rna/073304.119 – link to the publication
-
Molecular recognition of mRNA 5' cap by 3' poly(A)-specific ribonuclease (PARN) differs from interactions known for other cap-binding proteins
- Authors:
- Niedzwiecka A., Nilsson P., Worch R., Stepinski J., Darzynkiewicz E., Virtanen A.
- Academic press:
- Biochimica at Biophysica Acta - Proteins and Proteomics (year: 2016, volume: 1864, pages: 331-345), Publisher: Elsevier
- Status:
- Published
- DOI:
- 10.1016/j.bbapap.2016.01.002 – link to the publication
-
eIF4E-like complexes formed by cap-binding homolog TbEIF4E5 with TbEIF4G1 or TbEIF4G2 are implicated in post-transcriptional regulation in Trypanosoma brucei
- Authors:
- Freire E.R., Vashisht A.A., Malvezzi A.M., Zuberek J., Langousis G., Saada E.A., Nascimento J. DE F., Stepinski J., Darzynkiewicz E., Hill K., De Melo Neto O.P., Wohlschlegel J.A., Sturm N.R., Campbell D.A.
- Academic press:
- RNA (year: 2014, volume: 20, pages: 1272-1286), Publisher: Cold Spring Harbor Laboratory Press
- Status:
- Published
- DOI:
- 10.1261/rna.045534.114 – link to the publication
-
Decapping Scavenger (DcpS) enzyme: Advences in its structure, activity and roles in the cap-dependent mRNA metabolism
- Authors:
- Milac A.L., Bojarska E., Wypijewska del Nogal A.
- Academic press:
- Biochimica at Biophysica Acta - Gene Regulatory Mechanism (year: 2014, volume: 1839, pages: 452-462), Publisher: Elsevier
- Status:
- Published
- DOI:
- 10.1016/j.bbagrm.2014.04.007 – link to the publication
-
Helix 69 of Escherichia coli 23S ribosomal RNA as a peptide nucleic acid target
- Authors:
- Kulik M., Markowska-Zagrajek A., Wojciechowska M., Grzela R., Witula T., Trylska J.
- Academic press:
- Biochimie (year: 2017, volume: 138, pages: 32-42), Publisher: Elsevier
- Status:
- Published
- DOI:
- 10.1016/j.biochi.2017.04.001 – link to the publication
-
Quantum dots use both LUMO and surface trap electrons in photoreduction process
- Authors:
- Darzynkiewicz Z.M., Pedziwiatr M., Grzyb J.
- Academic press:
- Journal of Luminescence (year: 2017, volume: 183, pages: 401-409), Publisher: Elsevier
- Status:
- Published
- DOI:
- 10.1016/j.jlumin.2016.11.070 – link to the publication
-
Amino-Functionalized 5' Cap Analogs as Tools for Site-Specific Sequence-Independent Labeling of mRNA
- Authors:
- Warminski M., Sikorski P.J., Warminska Z., Lukaszewicz M., Kropiwnicka A., Zuberek J., Darzynkiewicz E., Kowalska J., Jemielity J.
- Academic press:
- Bioconjugate Chemistry (year: 2017, volume: 28, pages: 1978-1992), Publisher: American Chemical Society
- Status:
- Published
- DOI:
- 10.1021/acs.bioconjchem.7b00291 – link to the publication
-
Modified ARCA analogs providing enhanced translational properties of capped mRNAs
- Authors:
- Kocmik I., Piecyk K., Rudzinska M., Niedzwiecka A., Darzynkiewicz E., Grzela R., Jankowska-Anyszka M.
- Academic press:
- Cell Cycle (year: 2018, volume: 17, pages: 1624-1636), Publisher: Taylor & Francis
- Status:
- Published
- DOI:
- 10.1080/15384101.2018.1486164 – link to the publication
-
Hydrolytic activity of human Nudt16 enzyme on dinucleotide cap analogs and short capped oligonucleotides
- Authors:
- Grzela R., Nasilowska K., Lukaszewicz M., Tyras M., Stepinski J., Jankowska-Anyszka M., Bojarska E., Darzynkiewicz E.
- Academic press:
- RNA (year: 2018, volume: 24, pages: 633-642), Publisher: Cold Spring Harbor Laboratory Press
- Status:
- Published
- DOI:
- 10.1261/rna.065698.118 – link to the publication
-
Cap analgs modified with 1,2-dithiodiphosphate moiety protect mRNA from decapping and enhance its translational potential
- Authors:
- Strenkowska M., Grzela R., Majewski M., Wnek K., Kowalska J., Lukaszewicz M., Zuberek J., Darzynkiewicz E., Kuhn A.N., Sahin U., Jemielity J.
- Academic press:
- Nucleic Acids Research (year: 2016, volume: 44, pages: 9578-9590), Publisher: Oxford University Press
- Status:
- Published
- DOI:
- 10.1093/nar/gkw896 – link to the publication
-
Decapping Scavenger Enzyme Activity toward N2-substituted 5' end mRNA Cap Analogues
- Authors:
- Pietrow P., Ferenc-Mrozek A., Piecyk K., Bojarska E., Darzynkiewicz E., Jankowska-Anyszka M.
- Academic press:
- ACS Omega (year: 2019, volume: 4, pages: 17576-17580), Publisher: ACS Publications
- Status:
- Published
- DOI:
- 10.1021/acsomega.9b02715 – link to the publication
-
Cap analogs containing 6-thioguanosine - reagents for the synthesis of mRNAs selectively photo-crosslinkable with cap-binding biomolecules
- Authors:
- Nowakowska M., Kowalska J., Martin F., d'Orchymont A., Zuberek J., Lukaszewicz M., Darzynkiewicz E., Jemielity J.
- Academic press:
- Organic & Biomolecular Chemistry (year: 2014, volume: 12, pages: 4841-4847), Publisher: Royal Society of Chemistry
- Status:
- Published
- DOI:
- 10.1039/c4ob00059e – link to the publication
-
Effect of different N7 substitution of dinucleotide cap analogs on the hydrolytic susceptibility towards scavenger decapping enzymes (DcpS)
- Authors:
- Piecyk K., Darzynkiewicz Z.M., Jankowska-Anyszka M., Ferenc-Mrozek A., Stepinski J., Darzynkiewicz E., Bojarska E.
- Academic press:
- Biochemical and Biophysical Research Communications (year: 2015, volume: 464, pages: 89-93), Publisher: Elsevier
- Status:
- Published
- DOI:
- 10.1016/j.bbrc.2015.06.001 – link to the publication
-
Clickable trimethylguanosine cap analogs modified within the triphosphate bridge: synthesis, conjugation to RNA and susceptibility to degradation
- Authors:
- Wojtczak B.A., Warminski M., Kowalska J., Lukaszewicz M., Honcharenko M., Smith C.I.E., Stromberg R., Darzynkiewicz E., Jemielity J.
- Academic press:
- RCS Advances (year: 2016, volume: 6, pages: 8317-8328), Publisher: Royal Society of Chemistry
- Status:
- Published
- DOI:
- 10.1039/c5ra25684d – link to the publication
-
Phosphate-modified analogues of m7GTP and m7Gppppm7G - Synthesis and biochemical properties
- Authors:
- Ziemniak M., Kowalska J., Lukaszewicz M., Zuberek J., Wnek K., Darzynkiewicz E., Jemielity J.
- Academic press:
- Bioorganic & Medicinal Chemistry (year: 2015, volume: 23, pages: 5369-5381), Publisher: Elsevier
- Status:
- Published
- DOI:
- 10.1016/j.bmc.2015.07.052 – link to the publication
Book publications / chapters in book publications (1)
-
mRNA and snRNA Cap Analogs: Synthesis and Applications
- Authors:
- Stepinski J., Darzynkiewicz E.
- Book:
- Chemical Biology of Nucleic Acids - Fundamentals and Clinical Applications, RNA Technologies (year: 2014, volume: 28, pages: 511-561), Publisher: Springer-Verlag
- Status:
- Published