0000000000496790

AUTHOR

Joos Aschenbrenner

showing 2 related works from this author

Engineering of a DNA Polymerase for Direct m6A Sequencing

2017

Methods for the detection of RNA modifications are of fundamental importance for advancing epitranscriptomics. N6-methyladenosine (m6A) is the most abundant RNA modification in mammalian mRNA and is involved in the regulation of gene expression. Current detection techniques are laborious and rely on antibody-based enrichment of m6A-containing RNA prior to sequencing, since m6A modifications are generally "erased" during reverse transcription (RT). To overcome the drawbacks associated with indirect detection, we aimed to generate novel DNA polymerase variants for direct m6A sequencing. Therefore, we developed a screen to evolve an RT-active KlenTaq DNA polymerase variant that sets a mark for…

0301 basic medicineAdenosineRNA-dependent RNA polymeraseDNA-Directed DNA Polymerase010402 general chemistryProtein Engineering01 natural sciencesCatalysis03 medical and health sciencesDNA polymerasesSequencing by hybridization[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Genomics [q-bio.GN]TheoryofComputation_ANALYSISOFALGORITHMSANDPROBLEMCOMPLEXITYRNA polymerase IRNA MessengerPolymerasebiologyOligonucleotideN6-methyladenosineReverse Transcriptase Polymerase Chain ReactionCommunicationMultiple displacement amplificationHigh-Throughput Nucleotide Sequencing[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biologyGeneral ChemistryDNA MethylationRNA modificationMolecular biologyReverse transcriptaseCommunications0104 chemical sciencesSequencing by ligationenzyme engineering030104 developmental biologyComputingMethodologies_PATTERNRECOGNITIONddc:540biology.proteinepitranscriptomicsRNA Methylation
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Entwicklung einer DNA-Polymerase für die direkte m6A-Sequenzierung

2017

Methoden zur Analyse von RNA-Modifikationen sind essenziell fur das Forschungsfeld der Epitranskriptomik. N6-Methyladenosin (m6A) ist die haufigste Modifikation in der mRNA von Saugetieren und erfullt Funktionen in der Regulation der Genexpression. Techniken zur Detektion dieser Modifikation basieren derzeit auf der Anreicherung von m6A-haltigen RNA-Fragmenten durch Antikorper. Dieser m6A-spezifische Schritt vor der Sequenzierung ist notig, da die Information uber die Modifikation wahrend der reversen Transkription (RT) geloscht wird. Um die Nachteile einer solchen indirekten Detektion zu uberwinden, haben wir uns zum Ziel gesetzt, neue DNA-Polymerasen zu entwickeln, die eine direkte m6A-Se…

0301 basic medicine03 medical and health sciences030104 developmental biologyGeneral Medicine010402 general chemistry01 natural sciences0104 chemical sciencesAngewandte Chemie
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