DORQ-seq: high-throughput quantification of femtomol tRNA pools by combination of cDNA hybridization and Deep sequencing.

Due to its high modification content tRNAs are notoriously hard to quantify by reverse transcription and RNAseq. Bypassing numerous biases resulting from concatenation of enzymatic treatments, we here report a hybrid approach that harnesses the advantages of hybridization-based and deep sequencing-based approaches. The method renders obsolete any RNAseq related workarounds and correction factors that affect accuracy, sensitivity, and turnaround time.

Ischemic preconditioning: what else can it do…?

Ischemic preconditioning (IPC) can enhance maximal strength likely due to neural priming. Cruz et al. (Cruz R, Tramontin AF, Oliveira AS, Caputo F, Denadai BS, Greco CC.

N1-methylation of adenosine (mA) in ND5 mRNA leads to complex I dysfunction in Alzheimer's disease.

One mechanism of particular interest to regulate mRNA fate post-transcriptionally is mRNA modification. Especially the extent of mA mRNA methylation is highly discussed due to methodological differences. However, one single mA site in mitochondrial ND5 mRNA was unanimously reported by different groups.

Repeated-Sprint Training With Blood-Flow Restriction Improves Repeated-Sprint Ability Similarly to Unrestricted Training at Reduced External Loads.

Purpose: This study examined performance and physiological adaptations following 3 weeks of repeated-sprint training (RST) with blood-flow restriction (BFR) or without (non-BFR).

Methods: Twenty-six semiprofessional and amateur adult male team-sport players were assessed for repeated-sprint ability, anaerobic capacity, leg lean mass, neuromuscular function, and maximal aerobic capacity before and after RST. Participants completed 9 cycling RST sessions (3 sets of 5-7 × 5-s sprints, 25-s passive recovery, 3-min rest) over a 3-week period with BFR or non-BFR.

Manganese Ions Individually Alter the Reverse Transcription Signature of Modified Ribonucleosides.

Reverse transcription of RNA templates containing modified ribonucleosides transfers modification-related information as misincorporations, arrest or nucleotide skipping events to the newly synthesized cDNA strand. The frequency and proportion of these events, merged from all sequenced cDNAs, yield a so-called RT signature, characteristic for the respective RNA modification and reverse transcriptase (RT). While known for DNA polymerases in so-called error-prone PCR, testing of four different RTs by replacing Mg with Mn in reaction buffer revealed the immense influence of manganese chloride on derived RT signatures, with arrest rates on mA positions dropping from 82% down to 24%.

Graphical Workflow System for Modification Calling by Machine Learning of Reverse Transcription Signatures.

Modification mapping from cDNA data has become a tremendously important approach in epitranscriptomics. So-called reverse transcription signatures in cDNA contain information on the position and nature of their causative RNA modifications. Data mining of, e.