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Quillien, A.

Publications and source records attributed to Quillien, A..

4 recordsLinked to original sources

Prime-Editing in Marchantia paleacea: Expanding the Genome-Editing Toolbox in Bryophytes

Since the development of CRISPR-based genome editing tools, a number of novel technologies have emerged. This includes Prime-Editing that acts as a search and replace genome editing tool. Prime-Editing has been deployed across multiple clades, including in a few flowering plants. Here, we report on the development of an efficient Prime Editor (PE) for the model bryophyte Marchantia. Initial tests were conducted on Acetolactate Synthase as a target and revealed an average efficiency above 40%. The system has been developed in the GoldenGate cloning system, facilitating construct design. The development of PE in Marchantia expands the Genome-Editing tools available for this emerging model in plant biology.

plant biology↗

3D RNA profile of habenulae in the catshark Scyliorhinus canicula: a reference in jawed vertebrates

We report the generation of a spatially resolved RNA profile of the habenulae in a chondrichthyan, the catshark Scyliorhinus canicula. The digital profiles obtained for candidate markers of the broad territories previously characterized in this species reflect their broad characteristics, including left- or right asymmetric distributions. We use spatial autocorrelation and autocorrelation as measures to analyze these data and identify novel territory markers. These analyses lead to a refined molecular characterization of catshark habenulae and lead to extended lists of markers for the major territories previously identified. They confirm the marked asymmetry of the catshark habenulae, both in its lateral and medial components. In addition, they highlight a bilateral lateral to medial organization in radial bands, which may reflect a temporal regulation of neurogenesis. These data lead to a novel reference to explore the lmode of evolution of habenulae and of their asymmetries across gnathostomes.

developmental biology↗

Probing small ribosomal subunit RNA helix 45 acetylation across eukaryotic evolution

NAT10 is an essential enzyme that catalyzes the formation of N4-acetylcytidine (ac4C) in eukaryotic transfer RNA (tRNA) and 18S ribosomal RNA (rRNA). Recent studies in human cells suggested that rRNA acetylation is dependent on SNORD13, a non-canonical box C/D small nucleolar RNA (SNORD) predicted to base-pair with 18S rRNA via two antisense elements. However, the selectivity of SNORD13-dependent cytidine acetylation and its relationship to NAT10s essential function in pre-rRNA processing remain to be defined. Here, we used CRISPR-Cas9 genome editing to formally demonstrate that SNORD13 is required for acetylation of a single cytidine residue of human and zebrafish 18S rRNA. In-depth characterization revealed that SNORD13-dependent ac4C is dispensable for yeast or human cell growth, ribosome biogenesis, translation, and the development of multicellular metazoan model organisms. This loss of function analysis inspired a cross-evolutionary survey of the eukaryotic rRNA acetylation machinery that led to the characterization of many novel SNORD13 genes in phylogenetically-distant metazoans and more deeply rooted photosynthetic organisms. This includes an atypical SNORD13-like RNA in D. melanogaster which appears to guide ac4C to 18S rRNA helix 45 despite lacking one of the two rRNA antisense elements. Finally, we discover that C. elegans 18S rRNA is not acetylated despite the presence of an essential NAT10 homolog. Altogether, our findings shed light on the molecular mechanisms underlying SNORD13-mediated rRNA acetylation across the eukaryotic tree of life and raise new questions regarding the biological function and evolutionary persistence of this highly conserved rRNA base modification.

molecular biology↗

Prmt5 promotes vascular morphogenesis in zebrafish independently of its methyltransferase activity.

During development, the vertebrate vasculature undergoes major growth and remodeling. While the transcriptional cascade underlying blood vessel formation starts to be better characterized, little is known concerning the role and mode of action of epigenetic enzymes during this process. Here, we explored the role of the Protein Arginine Methyl Transferase Prmt5 during blood vessel formation and hematopoiesis in zebrafish. Through the generation of a prmt5 mutant, we highlighted a key role of Prmt5 in both hematopoiesis and blood vessel formation. Notably, we showed that Prmt5 promotes vascular morphogenesis through the transcriptional control of ETS transcription factors and adhesion proteins in endothelial cells. Interestingly, we found that Prmt5 methyltransferase activity is not required to regulate gene expression, and the comparison of chromatin architecture impact on reporter genes expression leads us to propose that Prmt5 rather regulates transcription by acting as a scaffold protein that facilitates chromatin looping in these cells.

developmental biology↗