Investigating RNA dynamics from single molecule transcriptomes.

Investigating RNA dynamics is crucial for uncovering fundamental mechanisms, such as alternative splicing, RNA stability, and post-transcriptional modifications, all processes with implications for identifying therapeutic targets and advancing knowledge of cellular function and regulation. Advances in long-read sequencing technologies, particularly from Pacific Biosciences (PacBio) and Oxford Nanopore Technologies (ONT), offer unprecedented insights into RNA dynamics at single molecule and single nucleotide resolutions. In this review, we examine protocols and methods for analyzing RNA dynamics, focusing on isoform detection, poly(A) tail length quantification, and mapping of RNA modifications.

Global mapping of BMAL1 protein-DNA interactions in human retinal Müller cells.

The circadian clock, a conserved biologic timekeeping mechanism, is pivotal in orchestrating rhythmic physiologic processes. While extensively studied in the central clock, the involvement of BMAL1 in peripheral clocks, particularly in human Müller cells, remains underexplored. Müller cells, critical for retinal homeostasis, may unveil novel insights into circadian regulation.

Nucleotide-resolution Mapping of RNA N6-Methyladenosine (m6A) modifications and comprehensive analysis of global polyadenylation events in mRNA 3' end processing in malaria pathogen .

is an obligate human parasite of the phylum Apicomplexa and is the causative agent of the most lethal form of human malaria. Although N6-methyladenosine modification is thought to be one of the major post-transcriptional regulatory mechanisms for stage-specific gene expression in apicomplexan parasites, the precise base position of m6A in mRNAs or noncoding RNAs in these parasites remains unknown. Here, we report global nucleotide-resolution mapping of m6A residues in using DART-seq technology, which quantitatively displayed a stage-specific, dynamic distribution pattern with enrichment near mRNA 3' ends.

Epitranscriptomics in parasitic protists: Role of RNA chemical modifications in posttranscriptional gene regulation.

"Epitranscriptomics" is the new RNA code that represents an ensemble of posttranscriptional RNA chemical modifications, which can precisely coordinate gene expression and biological processes. There are several RNA base modifications, such as N6-methyladenosine (m6A), 5-methylcytosine (m5C), and pseudouridine (Ψ), etc. that play pivotal roles in fine-tuning gene expression in almost all eukaryotes and emerging evidences suggest that parasitic protists are no exception.