Biology subjects

Moran, F.

Publications and source records attributed to Moran, F..

4 recordsLinked to original sources

bioRxiv2026

pH-dependent anti-TfR1 NANOBODY(R) molecules deliver efficacious oligonucleotide payloads to muscle and CNS tissues

The blood-brain barrier (BBB) is a highly selective, semi-permeable border of endothelial cells that prevents solutes and therapeutic agents in systemic circulation from passively crossing into the central nervous system (CNS) parenchyma. The Transferrin receptor 1 (TfR1) endocytosis pathway for iron homeostasis is one of the most well-characterized strategies for therapeutic delivery across the BBB. The work presented here showcases the discovery of novel anti-TfR1 NANOBODY(R) shuttles. The identified anti-TfR1 NANOBODY(R) molecules display cross-reactivity and pH-dependent binding to human, cynomolgus (cyno), and mouse TfR1. Structural data further explain and support the underlying mechanism of this pH-dependent binding. These anti-TfR1 NANOBODY(R) molecules were successfully conjugated to both short-interfering RNA (siRNA) and antisense oligonucleotide (ASO) tool payloads. anti-TfR1 NANOBODY(R)-siRNA conjugates can induce up to 60% knockdown of the target mRNA transcript in skeletal muscle up to two weeks post a single IV dose in mice and up to 35-40% at four weeks post dose. Furthermore, extending the half-life of the anti-TfR1 NANOBODY(R)-ASO shuttles enhances heart, sciatic nerve, and brain exposure and enables up to 30-60% target knockdown in different CNS cell types. Altogether, these results highlight important features for the development of anti-TfR1 shuttles for the purpose of downregulating target mRNA transcripts in muscle and CNS for a variety of neurologic and neuromuscular indications.

neuroscience↗

bioRxiv2026

A highly versatile real-time quantitative RT-PCR method and sampling strategies for the accurate detection of citrus yellow vein clearing virus

Citrus yellow vein clearing virus (CYVCV) is the causal agent of an emerging disease representing a potentially high-impact threat for citrus production. Despite remaining outside Europe for decades, CYVCV has now expanded towards two important European citrus producers, Italy and, more recently, Spain. The presence of this virus in the EPPO region represents a current threat with unpredictable and potentially devastating consequences for European citriculture. Therefore, urgent protective measures need to be taken to prevent CYVCV spread and minimize its impact. Diagnostics is a key measure in the management of viral diseases, highlighting the need for harmonized methods suitable for reliable routine detection of the currently known CYVCV diversity. In this study, an inclusive, efficient and highly sensitive real-time RT-qPCR for the detection of CYVCV in plant material and transmission vectors has been developed and validated according to EPPO standards. Moreover, the validated method has been successfully adapted to both PCR digital platforms, that allow high-sensitive absolute quantitative detection, essential in the diagnostics at low viral concentrations; and PCR portable tools, that can be applied in a real diagnostic context for on-site detection. This versatility combines standard validated performance, absolute sensitive quantitation and real on-site detection. The study has also addressed sampling strategies to support reliable molecular diagnostic performance. Our results represent an improvement in the detection of CYVCV to be applied in epidemiological studies and different real diagnostic contexts for the containment of this important citrus pathogen.

molecular biology↗

bioRxiv2026

High Throughput-based Surveillance Reveals New Components of Spanish Citrus Virome Related to Tristeza, Impietratura and Yellow Vein Clearing Diseases

Citrus is one of the most important crops cultivated worldwide, representing a strategic source of agricultural income for many countries, including Spain, the main producer within the European Union. The protection of this highly valuable industry against an increasing global movement of pests and pathogens requires effective regulatory measures, including control of plant propagation material, phytosanitary surveillance and risk assessment, which are based not only on knowledge of the established threats but also on potential emerging threats affecting citrus that may circulate unnoticed in the production system. In this work, with the aim of generating knowledge on potential emerging viruses in Spanish citrus orchards, high-throughput sequencing (HTS) analysis has been applied to monitor the sanitary status of several growing areas of one of the main citrus producer regions in Spain, the Valencian Community. The results of this study have revealed a much more complex citrus virome than previously reported, including citrus yellow vein clearing virus (CYVCV), a non-regulated but harmful citrus virus, as well as the T3 genotype of citrus tristeza virus (CTV) and citrus virus A (CiVA), not detected to date in Spain. Moreover, our results indicate the existence of other unknown components of the citrus virome. HTS detection of CYVCV, CTV T3 and CiVA and their presence in Spanish orchards has been confirmed by RT-PCR and Sanger sequencing. These findings have relevant implications in the development of control and regulatory measures against three important viral diseases, tristeza, impietratura and yellow vein clearing diseases, and demonstrate the added value of HTS-based surveillance to discover emerging components of the citrus virome.

molecular biology↗

bioRxiv2021

A two-level, dynamic fitness landscape of hepatitis C virus revealed by self-organized haplotype maps

Fitness landscapes reflect the adaptive potential of viruses. There is no information on how fitness peaks evolve when a virus replicates extensively in a controlled cell culture environment. Here we report the construction of Self-Organized Maps (SOMs), based on deep sequencing reads of three amplicons of the NS5A-NS5B-coding region of hepatitis C virus (HCV). A two-dimensional neural network was constructed and organized according to sequence relatedness. The third dimension of the fitness profile was given by the haplotype frequencies at each neuron. Fitness maps were derived for 44 HCV populations that share a common ancestor that was passaged up to 210 times in human hepatoma Huh-7.5 cells. As the virus increased its adaptation to the cells, the number of fitness peaks expanded, and their distribution shifted in sequence space. The landscape consisted of an extended basal platform, and a lower number of protruding higher fitness peaks. The function that relates fitness level and peak abundance corresponds a power law, a relationship observed with other complex natural phenomena. The dense basal platform may serve as spring-board to attain high fitness peaks. The study documents a highly dynamic, double-layer fitness landscape of HCV when evolving in a monotonous cell culture environment. This information may help interpreting HCV fitness landscapes in complex in vivo environments. IMPORTANCEThe study provides for the first time the fitness landscape of a virus in the course of its adaptation to a cell culture environment, in absence of external selective constraints. The deep sequencing-based self-organized maps document a two-layer fitness distribution with an ample basal platform, and a lower number of protruding, high fitness peaks. This landscape structure offers potential benefits for virus resilience to mutational inputs.

microbiology↗