RESUMO
A gold nanogap substrate was used to measure the thiram and carbaryl residues in various fruit juices using surface-enhanced Raman scattering (SERS). The gold nanogap substrates can detect carbaryl and thiram with limits of detection of 0.13 ppb (0.13 µgkg-1) and 0.22 ppb (0.22 µgkg-1). Raw SERS data were first preprocessed to reduce noise and undesirable effects and, were later used for model creation, implementing classification, and regression analysis techniques. The partial least-squares regression models achieved the highest prediction correlation coefficient (R2) of 0.99 and the lowest root mean square of prediction value below 0.62 ppb for both pesticide-infected juice samples. Furthermore, to differentiate between juice samples contaminated by both pesticides and control (pesticide-free), logistic-regression classification models were produced and achieved the highest classification accuracies of 100% and 99% for contaminated juice containing thiram and 100% accurate results for contaminated juice containing carbaryl. This indicates that the gold nanogap surface has significant potential for achieving high sensitivity in detecting trace contaminants in food samples.
Assuntos
Carbaril , Contaminação de Alimentos , Sucos de Frutas e Vegetais , Ouro , Resíduos de Praguicidas , Análise Espectral Raman , Tiram , Resíduos de Praguicidas/análise , Análise Espectral Raman/métodos , Carbaril/análise , Sucos de Frutas e Vegetais/análise , Tiram/análise , Contaminação de Alimentos/análise , Ouro/química , Quimiometria , Nanopartículas Metálicas/química , Limite de Detecção , Frutas/químicaRESUMO
This study reported an application of Au nanogap substrates for surface-enhanced Raman scattering (SERS) measurements to quantitatively analyze melamine and its derivative products at trace levels in pet liquid food (milk) combined with a waveband selection approach, namely variable importance in projection (VIP). Six different concentrations of melamine, cyanuric acid, and melamine combined with cyanuric acid were created, and SERS spectra were acquired from 550 to 1620cm-1. Detection was possible up to 200 pM for melamine-contaminated samples, and 400 pM concentration detection for other two groups. The VIP-PLSR models obtained correlation coefficient (R2) values of 0.997, 0.985, and 0.981, with root mean square error of prediction (RMSEP) values of 18.492 pM, 19.777 pM, and 15.124 pM for prediction datasets. Additionally, partial least square discriminant analysis (PLS-DA) was used to classify both pure and different concentrations of spiked samples. The results showed that the maximum classification accuracy for melamine was 100%, for cyanuric acid it was 96%, and for melamine coupled with cyanuric acid it was 95%. The results obtained clearly demonstrated that the Au nanogap substrate offers low-concentration, rapid, and efficient detection of hazardous additive chemicals in pet consuming liquid food.
RESUMO
Molybdenum sulfide-oxide (MoS2, MS) emerges as the prime electrocatalyst candidate demonstrating hydrogen evolution reaction (HER) activity comparable to platinum (Pt). This study presents a facile electrochemical approach for fabricating a hybrid copper (Cu)/MoS2 (CMS) nanostructure thin-film electrocatalyst directly onto nickel foam (NF) without a binder or template. The synthesized CMS nanostructures were characterized utilizing energy-dispersive X-ray spectroscopy (EDS), scanning electron microscopy (SEM), X-ray diffraction (XRD), and electrochemical methods. The XRD result revealed that the Cu metal coating on MS results in the creation of an extremely crystalline CMS nanostructure with a well-defined interface. The hybrid nanostructures demonstrated higher hydrogen production, attributed to the synergistic interplay of morphology and electron distribution at the interface. The nanostructures displayed a significantly low overpotential of -149 mV at 10 mA cm-2 and a Tafel slope of 117 mV dec-1, indicating enhanced catalytic activity compared to pristine MoS2.This research underscores the significant enhancement of the HER performance and conductivity achieved by CMS, showcasing its potential applications in renewable energy.
RESUMO
Background: Suicidal thoughts and behaviors (STBs) are prevalent within the Lesbian, Gay, Bisexual, Transgender, and Queers (LGBTQ) community, often exacerbated by challenges in accessing care and the perceived stigma and discrimination tied to disclosing one's identity. Digital health interventions that offer psychosocial self-help present a promising platform to reach individuals at risk of STBs, especially those who may not engage with conventional health services. This review aimed to assess the role of digital-based intervention in reducing STBs among LGBTQ individuals. Methods: We conducted a systematic literature search from three databases, PsycINFO, PubMed, and CINHAL, from 1st Jan 1990 to 31st December 2023. The review encompassed studies investigating the feasibility, acceptability, and impact of digital interventions on STBs, employing randomized control trials (RCTs), pseudo-RCTs, observational pre-posttest designs, and qualitative studies. Potential bias was evaluated using the McGill Mixed Methods Appraisal Tool (MMAT). Results: Five non-overlapping studies were included, reporting data from 777 participants. The studies featured diverse types of digital interventions, including videos, online writing, and mobile applications. The studies included three RCTs, and two qualitative studies. Across most of these studies, notable enhancements or reductions in the proportion of participants reporting STBs were observed post-intervention, alongside improvements in help-seeking intentions. The findings underscored that the applications used in the studies were engaging, acceptable, and deemed feasible in effectively addressing suicide prevention among the LGBTQ community. Conclusion: Overall, digital interventions were found to be feasible and acceptable in suicide prevention among LGBTQ communities, demonstrating preliminary efficacy in increasing help-seeking behavior when experiencing suicidal thoughts and in reducing STBs. Therefore, advocating for widespread promotion and dissemination of digital health interventions is crucial, particularly in low- and middle-income countries (LMICs) with limited access to health services and heightened barriers to obtaining such services. Further research using fully powered RCT is imperative to assess the efficacy of these interventions.
RESUMO
Non-invasive and passive detection of explosives in the vapor phase is advantageous for military, counter-terrorism, and homeland security applications. Detection of explosives using SERS has been an active research topic. However, the vapor pressures of most explosives are low at room temperature, and consequently, the vapor phase detection by SERS is highly challenging without intentionally heating explosive powder to increase the vapor pressure. In this work, we report the rapid and sensitive detection of 2,4,6-trinitrotoluene (TNT) and 2,4-dinitrotoluene (2,4-DNT) in the vapor phase, using a gold nanogap (AuNG) SERS substrate. The AuNG SERS substrate was fabricated with electron beam evaporation, rapid thermal annealing, and wet etching. SERS measurements were carried out with an incident power as low as 0.56 mW at 785 nm. To prevent the condensation effect, the TNT and 2,4-DNT powders inside the cuvette were taken out before inserting the nanogap substrate. Our SERS results demonstrate the feasibility of the non-invasive detection of vapor phase explosives under ambient conditions.
RESUMO
Plasmonic molecules, which are geometrically well-defined plasmonic metal nanoparticle clusters, have attracted significant attention due to their enhancement of light-matter interactions owing to a stronger electric field enhancement than that by single particles. High-resolution lithography techniques provide precise positioning of plasmonic nanoparticles, but their fabrication costs are excessively high. In this study, we propose a lithography-free, self-assembly fabrication method, termed the dual-dewetting process, which allows the control of the size and density of gold nanoparticles. This process involves depositing a gold thin film on a substrate and inducing dewetting through thermal annealing, followed by a second deposition and annealing. The method achieves a uniform distribution of particle size and density, along with increased particle density, across a 6-inch wafer. The superiority of the method is confirmed by a 30-fold increase in the signal intensity of surface-enhanced Raman scattering following the additional dewetting with an 8 nm film, compared to single dewetting alone. Our findings indicate that the dual-dewetting method provides a simple and efficient approach to enable a variety of plasmonic applications through efficient plasmonic molecule large-area fabrication.
Assuntos
Nanopartículas Metálicas , Ouro , Eletricidade , Filmes Cinematográficos , Tamanho da PartículaRESUMO
Conventional spectroscopic methods like IR, and Raman are not very effective at detecting low levels of pesticides or harmful chemicals in food matrices. A quick, highly accurate approach that can identify pesticides present in different food products at lower levels must be developed in order to address this problem and ensure food safety. In this study, a highly sensitive and uniform wafer-scale Au nanogap surface-enhanced Raman spectroscopy (SERS) substrate was used for the quantitative analysis of carbaryl pesticide levels in standard solution, mango juice, and milk samples using chemometrics. Carbaryl was detected up to 3 ppb concentration levels for all three group of samples. However, due to the higher sensitivity, uniformity, and enhancement factors of the SERS substrate used in this investigation, the limit of detection (LOD) values for the standard solution, mango juice, and milk were 0.37 ppb, 0.57 ppb, and 0.15 ppb at 1380 cm-1, 1380 cm-1, and 1364 cm-1 wavenumber ranges. In order to predict different carbaryl concentrations (1, 2, 3, 4, and 5 ppb), the variable importance in projection (VIP) method combined with partial least squares regression (PLSR) and attained the coefficient of determination (R2) values of 0.994, 0.989, and 0.978 along with minimum root mean square error (RMSE) values of 0.112, 0.190, and 0.278 ppb for the prediction datasets. Furthermore, PLS-DA was able to distinguish between pure and adulterated samples with the highest classification accuracy of 100 % for a standard solution, and mango juice and 94.4 % for milk samples. Considering this, we can conclude that the SERS Au Nanogap substrate can rapidly and effectively detect carbaryl pesticides quantitatively and qualitatively in mango juice and milk.
Assuntos
Praguicidas , Animais , Praguicidas/análise , Carbaril/análise , Leite/química , Análise Espectral Raman/métodos , Inocuidade dos Alimentos , Ouro/químicaRESUMO
BACKGROUND: In 2021, the Nepal national emergency care system's assessment (ECSA) identified 39 activities and 11 facility-specific goals to improve care. To support implementation of the ECSA facility-based goals, this pilot study used the World Health Organization's (WHO) Hospital Emergency Unit Assessment Tool (HEAT) to evaluate key functions of emergency care at tertiary hospitals in Kathmandu, Nepal. METHODS: This cross-sectional study used the standardized HEAT assessment tool. Data on facility characteristics, human resources, clinical services, and signal functions were gathered via key informant interviews conducted by trained study personnel. Seven tertiary referral centers in the Kathmandu valley were selected for pilot evaluation including governmental, academic, and private hospitals. Descriptive statistics were generated, and comparative analyses were conducted. RESULTS: All facilities had continuous emergency care services but differed in the extent of availability of each item surveyed. Academic institutions had the highest rating with greater availability of consulting services and capacity to perform specific signal functions including breathing interventions and sepsis care. Private institutions had the highest infrastructure availability and diagnostic testing capacity. Across all facilities, common barriers included lack of training of key emergency procedures, written protocols, point-of-care testing, and ancillary patient services. CONCLUSION: This pilot assessment demonstrates that the current emergency care capacity at representative tertiary referral hospitals in Kathmandu, Nepal is variable with some consistent barriers which preclude meeting the ECSA goals. The results can be used to inform emergency care development within Nepal and demonstrate that the WHO HEAT assessment is feasible and may be instructive in systematically advancing emergency care delivery at the national level if implemented more broadly.
RESUMO
Both rabies and snakebite primarily affect underserved and impoverished communities globally, with an estimated 200,000 people dying from these diseases annually, and the greatest burden being in Africa and Asia. Both diseases have been neglected and have thus been denied appropriate prioritization, support, and interventions, and face many of the challenges common to all neglected tropical diseases (NTDs). In line with the call for integrated approaches between NTDs in the recent NTD Roadmap, we sought to build upon previous conceptualizations for an integrated approach by identifying the commonalities between snakebite and rabies to explore the feasibility of an integrated approach. While multiple areas for potential integration are identified, we highlight the potential pitfalls to integrating rabies and snakebite programs, considering the nuances that make each disease and its intervention program unique. We conclude that health system strengthening, and capacity building should be the focus of any integrated approach among NTDs, and that by strengthening overall health systems, both rabies and snakebite can advocate for further support from governments and stakeholders.
RESUMO
Detecting trace amounts of explosives to ensure personal safety is important, and this is possible by using laser-based spectroscopy techniques. We performed surface-enhanced Raman scattering (SERS) using plasmonic nanogap substrates for the solution phase detection of some nitro-based compounds, taking advantage of the hot spot at the nanogap. An excitation wavelength of 785 nm with an incident power of as low as ≈0.1 mW was used to excite the nanogap substrates. Since both RDX and PETN cannot be dissolved in water, acetone was used as a solvent. TNT was dissolved in water as well as in hexane. The main SERS peaks of TNT, RDX, and PETN were clearly observed down to the order of picomolar concentration. The variations in SERS spectra observed from different explosives can be useful in distinguishing and identifying different nitro-based compounds. This result indicates that our nanogap substrates offer an effective approach for explosives identification.
RESUMO
The N6-methyladenosine (m6 A) RNA modification serves crucial functions in RNA metabolism; however, the molecular mechanisms underlying the regulation of m6 A are not well understood. Here, we establish arginine methylation of METTL14, a component of the m6 A methyltransferase complex, as a novel pathway that controls m6 A deposition in mammalian cells. Specifically, protein arginine methyltransferase 1 (PRMT1) interacts with, and methylates the intrinsically disordered C terminus of METTL14, which promotes its interaction with RNA substrates, enhances its RNA methylation activity, and is crucial for its interaction with RNA polymerase II (RNAPII). Mouse embryonic stem cells (mESCs) expressing arginine methylation-deficient METTL14 exhibit significantly reduced global m6 A levels. Transcriptome-wide m6 A analysis identified 1,701 METTL14 arginine methylation-dependent m6 A sites located in 1,290 genes involved in various cellular processes, including stem cell maintenance and DNA repair. These arginine methylation-dependent m6 A sites are associated with enhanced translation of genes essential for the repair of DNA interstrand crosslinks; thus, METTL14 arginine methylation-deficient mESCs are hypersensitive to DNA crosslinking agents. Collectively, these findings reveal important aspects of m6 A regulation and new functions of arginine methylation in RNA metabolism.
Assuntos
Adenosina/análogos & derivados , Arginina/química , Metiltransferases/metabolismo , Células-Tronco Embrionárias Murinas/metabolismo , Processamento de Proteína Pós-Traducional , Proteína-Arginina N-Metiltransferases/metabolismo , RNA Polimerase II/metabolismo , Adenosina/química , Animais , Citoplasma , Metiltransferases/química , Metiltransferases/genética , Camundongos , Células-Tronco Embrionárias Murinas/citologia , Proteína-Arginina N-Metiltransferases/genética , RNA Polimerase II/genética , TranscriptomaRESUMO
Highly efficient nanoparticle-on-metallic-mirror (NPOM) systems with a large gap size exhibiting good plasmonic enhancement are desirable for numerous practical applications. Careful, explicit design optimization strategies are required for preparing NPOMs and it is especially important in utilizing spherical nanoparticles. In this work, a new design blueprint for evaluating the role of random facets in spherical nanoparticles was investigated in detail to realize optimal NPOMs. We found that a precise single facet positioned at the nanoparticle's cavity outperformed multiple random facets due to the gap mode contribution. Differences and changes in the plasmonic modes were interpreted with the help of three-dimensional surface charge density mappings. A high-performance, single, bottom-faceted NPOM device with a large gap size (example 20 nm) was realized having 80-50% facet design, resulting in excellent gap mode enhancement. We succeeded in fabricating single bottom-faceted NPOMs (the non-facet region had a smooth spherical surface) with a large-scale unidirectionality (2 cm × 1.5 cm). Simulations and experimental characterizations of these components displayed excellent agreement. Our highly efficient NPOM design with a large gap size(s) enables interesting practical applications in the field of quantum emitters, energy devices, fuel generation and plasmon chemistry.
RESUMO
BACKGROUND: Sexual harassment has received global attention and has been recognized as a public health problem with increased physical and mental health risks among the victims. This study attempted to assess understanding, experience, response and effect of sexual harassment among the secondary school female students in Kathmandu, Nepal. METHODS: It was a mixed method study conducted among 441 secondary level female students. In quantitative component, a structured questionnaire was used to collect information relating to socio-demographic characteristics, and understanding, experience, response and effect of sexual harassment from 441 participants from three clusters (schools) of Tokha Municipality, Kathmandu. For qualitative method, two focused group discussions were conducted to assess the understanding of and response to sexual harassment. Quantitative and qualitative data were analyzed using Chi square test and thematic analysis method respectively. RESULTS: Around 76% of the participants had experienced some forms of sexual harassment in their life. Sexual harassment was significantly associated with religion and parental occupation. Majority of participants who faced sexual harassment reported that the perpetrators were strangers and they either ignored it or did nothing at the time of events. School, roads and public places were the most common settings where participants encountered sexual harassment. CONCLUSIONS: Sexual harassment is prevalent among female secondary students in Kathmandu. However, they lacked a clear understating of and ways to respond to sexual harassment. Raising awareness to recognize and respond to different forms of harassment and to change the victim-blaming attitudes can be a potential strategy for tackling the problem in Nepalese society.
Assuntos
Assédio Sexual/psicologia , Assédio Sexual/estatística & dados numéricos , Estudantes/psicologia , Adolescente , Feminino , Grupos Focais , Humanos , Nepal/epidemiologia , Prevalência , Religião , Instituições Acadêmicas/estatística & dados numéricos , Fatores Socioeconômicos , Adulto JovemRESUMO
A major limitation of RNA sequencing (RNA-seq) analysis of alternative splicing is its reliance on high sequencing coverage. We report DARTS (https://github.com/Xinglab/DARTS), a computational framework that integrates deep-learning-based predictions with empirical RNA-seq evidence to infer differential alternative splicing between biological samples. DARTS leverages public RNA-seq big data to provide a knowledge base of splicing regulation via deep learning, thereby helping researchers better characterize alternative splicing using RNA-seq datasets even with modest coverage.
Assuntos
Aprendizado Profundo , Splicing de RNA , RNA/análise , Análise de Sequência de RNA , Algoritmos , Processamento Alternativo , Teorema de Bayes , Epigenômica , Éxons , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Células Hep G2 , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Células K562 , Modelos Estatísticos , RNA/genética , Reprodutibilidade dos Testes , Processamento de Sinais Assistido por ComputadorRESUMO
5-methylcytosine (m5C) is a post-transcriptional RNA modification identified in both stable and highly abundant tRNAs and rRNAs, and in mRNAs. However, its regulatory role in mRNA metabolism is still largely unknown. Here, we reveal that m5C modiï¬cation is enriched in CG-rich regions and in regions immediately downstream of translation initiation sites and has conserved, tissue-specific and dynamic features across mammalian transcriptomes. Moreover, m5C formation in mRNAs is mainly catalyzed by the RNA methyltransferase NSUN2, and m5C is specifically recognized by the mRNA export adaptor ALYREF as shown by in vitro and in vivo studies. NSUN2 modulates ALYREF's nuclear-cytoplasmic shuttling, RNA-binding affinity and associated mRNA export. Dysregulation of ALYREF-mediated mRNA export upon NSUN2 depletion could be restored by reconstitution of wild-type but not methyltransferase-defective NSUN2. Our study provides comprehensive m5C profiles of mammalian transcriptomes and suggests an essential role for m5C modification in mRNA export and post-transcriptional regulation.
Assuntos
5-Metilcitosina/metabolismo , Metiltransferases/metabolismo , Proteínas Nucleares/metabolismo , Transporte de RNA/genética , Proteínas de Ligação a RNA/metabolismo , Fatores de Transcrição/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Células HeLa , Humanos , Masculino , Modelos Biológicos , Proteínas Nucleares/química , Especificidade de Órgãos/genética , Ligação Proteica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/química , Testículo/embriologia , Testículo/metabolismo , Fatores de Transcrição/químicaRESUMO
Among myriads of distinct chemical modifications in RNAs, dynamic N6-methyladenosine (m(6)A) is one of the most prevalent modifications in eukaryotic mRNAs and non-coding RNAs. Similar to the critical role of chemical modifications in regulation of DNA and protein activities, RNA m(6)A modification is also observed to be involved in the regulation of diverse functions of RNAs including meiosis, fertility, development, cell reprogramming and circadian period. The RNA m(6)A modification is recognized by YTH domain containing family proteins comprising of YTHDC1-2 and YTHDF1-3. Here we focus on the nuclear m(6)A reader YTHDC1 and its regulatory role in alternative splicing and other RNA metabolic processes.
Assuntos
Adenosina/análogos & derivados , Splicing de RNA , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Adenosina/metabolismo , Processamento Alternativo , Animais , Humanos , Metilação , Ligação Proteica , Fatores de Processamento de RNA/metabolismo , Fatores de Processamento de Serina-Arginina/metabolismo , Transdução de SinaisRESUMO
The regulatory role of N(6)-methyladenosine (m(6)A) and its nuclear binding protein YTHDC1 in pre-mRNA splicing remains an enigma. Here we show that YTHDC1 promotes exon inclusion in targeted mRNAs through recruiting pre-mRNA splicing factor SRSF3 (SRp20) while blocking SRSF10 (SRp38) mRNA binding. Transcriptome assay with PAR-CLIP-seq analysis revealed that YTHDC1-regulated exon-inclusion patterns were similar to those of SRSF3 but opposite of SRSF10. In vitro pull-down assay illustrated a competitive binding of SRSF3 and SRSF10 to YTHDC1. Moreover, YTHDC1 facilitates SRSF3 but represses SRSF10 in their nuclear speckle localization, RNA-binding affinity, and associated splicing events, dysregulation of which, as the result of YTHDC1 depletion, can be restored by reconstitution with wild-type, but not m(6)A-binding-defective, YTHDC1. Our findings provide the direct evidence that m(6)A reader YTHDC1 regulates mRNA splicing through recruiting and modulating pre-mRNA splicing factors for their access to the binding regions of targeted mRNAs.
Assuntos
Proteínas de Ciclo Celular/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Splicing de RNA , Proteínas de Ligação a RNA/metabolismo , Proteínas Repressoras/metabolismo , Adenosina/análogos & derivados , Adenosina/metabolismo , Sítios de Ligação , Éxons , Células HeLa , Humanos , Fatores de Processamento de RNA , RNA Mensageiro/metabolismo , Fatores de Processamento de Serina-ArgininaRESUMO
DNA double-strand breaks (DSBs) are highly cytotoxic lesions and pose a major threat to genome stability if not properly repaired. We and others have previously shown that a class of DSB-induced small RNAs (diRNAs) is produced from sequences around DSB sites. DiRNAs are associated with Argonaute (Ago) proteins and play an important role in DSB repair, though the mechanism through which they act remains unclear. Here, we report that the role of diRNAs in DSB repair is restricted to repair by homologous recombination (HR) and that it specifically relies on the effector protein Ago2 in mammalian cells. Interestingly, we show that Ago2 forms a complex with Rad51 and that the interaction is enhanced in cells treated with ionizing radiation. We demonstrate that Rad51 accumulation at DSB sites and HR repair depend on catalytic activity and small RNA-binding capability of Ago2. In contrast, DSB resection as well as RPA and Mre11 loading is unaffected by Ago2 or Dicer depletion, suggesting that Ago2 very likely functions directly in mediating Rad51 accumulation at DSBs. Taken together, our findings suggest that guided by diRNAs, Ago2 can promote Rad51 recruitment and/or retention at DSBs to facilitate repair by HR.
Assuntos
Proteínas Argonautas/metabolismo , Quebras de DNA de Cadeia Dupla , Reparo do DNA , Recombinação Homóloga , Rad51 Recombinase/metabolismo , Linhagem Celular , Humanos , Mapas de Interação de Proteínas , RNA/metabolismo , Ribonuclease III/metabolismoRESUMO
The methyltransferase like 3 (METTL3)-containing methyltransferase complex catalyzes the N6-methyladenosine (m6A) formation, a novel epitranscriptomic marker; however, the nature of this complex remains largely unknown. Here we report two new components of the human m6A methyltransferase complex, Wilms' tumor 1-associating protein (WTAP) and methyltransferase like 14 (METTL14). WTAP interacts with METTL3 and METTL14, and is required for their localization into nuclear speckles enriched with pre-mRNA processing factors and for catalytic activity of the m6A methyltransferase in vivo. The majority of RNAs bound by WTAP and METTL3 in vivo represent mRNAs containing the consensus m6A motif. In the absence of WTAP, the RNA-binding capability of METTL3 is strongly reduced, suggesting that WTAP may function to regulate recruitment of the m6A methyltransferase complex to mRNA targets. Furthermore, transcriptomic analyses in combination with photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation (PAR-CLIP) illustrate that WTAP and METTL3 regulate expression and alternative splicing of genes involved in transcription and RNA processing. Morpholino-mediated knockdown targeting WTAP and/or METTL3 in zebrafish embryos caused tissue differentiation defects and increased apoptosis. These findings provide strong evidence that WTAP may function as a regulatory subunit in the m6A methyltransferase complex and play a critical role in epitranscriptomic regulation of RNA metabolism.