🧬 PubMed RNA Editing Daily Digest

Listeria monocytogenes is an important foodborne pathogen associated with high mortality rates, especially among vulnerable populations, and therefore requires diagnostic methods that are not only highly sensitive and rapid but also suitable for use in resource-limited settings. In this study, we developed an isothermal amplification assay integrated with a lateral flow test (LFT) for reliable detection of L.monocytogenes. Two assay formats were designed and compared: (1) loop-mediated isothermal amplification (LAMP) with LFT detection of fluorescein- and biotin-labeled amplicons, and (2) LAMP combined with CRISPR/Cas12a, using LFT to detect a cleaved fork-shaped enhanced probe labeled with three fluoresceins. Both LFT formats utilized a common conjugate of gold nanoparticles and anti-fluorescein antibodies (anti-FAM), but differed in the test zone immobilization strategy: streptavidin for LAMP, and anti-FAM for LAMP-CRISPR/Cas12a. Among 12 tested (primer - label) combinations, the most effective was identified, but the sensitivity of the LAMP-LFT format was limited by high signal variability. In contrast, the LAMP-CRISPR/Cas12a assay, targeting LAMP amplicons with guide RNA, achieved a detection limit of 0.9 copies/reaction-representing > 20,000-fold improvement in detectable DNA concentration compared with LAMP-LFT-and comparable to fluorescence-based detection techniques. The LAMP-CRISPR/Cas12a-LFT assay was first reported to detect L.monocytogenes cells following thermal lysis (10 min at 95 °C), with a single-cell detection limit (0.2 cells/reaction in buffer, 1 cells/reaction in spiked milk) and an analysis time of 80 min. These results demonstrate the potential of the approach for sensitive, equipment-free detection of foodborne pathogens in complex food matrices.

Skin cancer is the third most common malignancy, with melanoma being the most challenging due to its resistance to current therapies. Gene editing technologies like CRISPR/Cas9 offer a promising strategy for targeting cancer-specific genes, but the efficient delivery of these tools to tumor sites remains a significant challenge. Lipid nanoparticles (LNPs) have emerged as the leading platform for gene editing tools due to their ability to protect and transport large payloads. To enhance the precision of gene editing in melanoma, we developed CD44-specific peptide-modified LNPs for targeted delivery of CRISPR/Cas9 mRNA and guide RNA against polo-like kinase 1 (sgPLK1). Our approach led to enhanced targeting and gene editing efficacy by specifically delivering CRISPR/Cas9 and sgPLK1 to melanoma tumor cells, resulting in significant inhibition of tumor growth in both in vitro and in vivo skin melanoma models. Moreover, this platform showed the capacity to reach metastatic melanoma in the brain and resulting in substantial suppression of tumor growth in brain metastasis models. We envision that this peptide-modification strategy could be further employed to improve the targeting capabilities and therapeutic outcomes of LNPs for CRISPR/Cas9-based gene editing, paving the way for more precise and effective cancer treatments.

Post-transcriptional regulation of AMPA-type ionotropic glutamate receptors (AMPARs) emerged in early vertebrates alongside a major expansion of AMPAR auxiliary subunits. Recent work shows that two key modifications-alternative splicing of the flip/flop cassette and Q/R site RNA editing-fine-tune channel gating and Ca

Bovine parainfluenza virus type 3 (BPIV3) is a major respiratory pathogen in cattle, with a substantial economic impact on the global livestock industry. The V protein, a nonstructural accessory protein expressed via RNA editing of the P gene, antagonizes canonical antiviral signaling. However, its additional role in promoting viral replication remains poorly defined. Here, we demonstrate that the BPIV3 V protein significantly enhances viral replication via a novel autophagy-dependent mechanism. Using a reverse genetics system, we generated a V-gene deletion mutant (rBPIV3-ΔV). Comparative transcriptomic (RNA-seq) analysis of cells infected with wild-type BPIV3 and rBPIV3-ΔV revealed that the V protein specifically modulates host autophagy signaling. We confirmed key differentially expressed genes using real-time quantitative PCR and identified the V protein as the essential viral factor driving BPIV3-induced autophagy. Collectively, our transcriptomic data delineate the molecular differences underlying the attenuated replication of rBPIV3-ΔV and establish that the V protein exploits the autophagy pathway to facilitate viral propagation in vitro. This finding provides a crucial theoretical advance in understanding BPIV3 pathogenesis and reveals potential targets for the development of novel antivirals and vaccines.