GenePub Date : 2024-11-05DOI: 10.1016/j.gene.2024.149080
Yinan Zhou, Ying Zhang, Dongchen Xu, Chenguang Yang, Xiao Lin, Kunlin Jin, Lei Xia, Qichuan Zhuge, Su Yang
{"title":"Exosomes from polarized Microglia: Proteomic insights into potential mechanisms affecting intracerebral hemorrhage.","authors":"Yinan Zhou, Ying Zhang, Dongchen Xu, Chenguang Yang, Xiao Lin, Kunlin Jin, Lei Xia, Qichuan Zhuge, Su Yang","doi":"10.1016/j.gene.2024.149080","DOIUrl":"https://doi.org/10.1016/j.gene.2024.149080","url":null,"abstract":"<p><p>Intracerebral hemorrhage (ICH) is a devastating form of stroke associated with significant morbidity and mortality. Microglia are intracranial innate immune cell that play critical roles in Intracerebral hemorrhage through direct or indirect means. Vesicle transport is a fundamental mechanism of intercellular communication. Recent studies have identified microglia in specific polarized states correlate with pathogenesis, material and signal transmission in ICH through derived extracellular vesicles. Diverse polarization states trigger distinct functions, however, the exosome proteomes across these states remain poorly characterized. Here, we hypothesized that microglia exosomal profiles vary with polarization states, impacting their functional repertoire and influencing outcomes in cerebral hemorrhage. In vitro model of cerebral hemorrhage, administration of 20 μg/ml LPS-induced M1 microglia derived exosomes (M1-Exo) with HT22 enhanced hemin-induced neuronal death, while IL-4-induced M2 microglia derived exosomes (M2-Exo) significantly reduced hemin-induced cell apoptosis and inflammation. Then we identified novel state-specific proteomic profiles of microglia-derived exosomes under these polarization conditions through label-free quantitative mass spectrometry (LFQ-MS). Analysis of protein content identified several exosomal signature proteins and hundreds of differentially expressed proteins across polarization states. Specifically, proteins including UMOD, NLRP3, ACOD1, IL1RN, heme oxygenase 1 (HMOX1), CCL4, and TNFRSF1B in M1-Exo were enriched in inflammatory pathways, while those in M2-Exo exhibited enrichment in autophagy, ubiquitination, and mitochondrial respiration. The analysis of those diverse exosomal proteins suggested unique proteomic profiles and possible intracellular signal transmission and regulation mechanisms. Together, these findings offer new insights and resources for studying microglia-derived exosome and pave the way for the development of novel therapeutic strategies targeting microglial exosome-mediated pathways.</p>","PeriodicalId":12499,"journal":{"name":"Gene","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142603974","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
GenePub Date : 2024-11-05DOI: 10.1016/j.gene.2024.149079
Modibo Diallo, Alicia Defay-Stinat, Victor Gindensperger, Angèle Sequeira, Aurélien Trimouille, Sophie Javerzat, Laetitia Bourgeade, Claudio Plaisant, Eulalie Lasseaux, Vincent Michaud, Isabelle Drumare, Benoit Arveiler
{"title":"A 65 kilobase deletion of the upstream TYR gene region in a family with oculocutaneous albinism type 1.","authors":"Modibo Diallo, Alicia Defay-Stinat, Victor Gindensperger, Angèle Sequeira, Aurélien Trimouille, Sophie Javerzat, Laetitia Bourgeade, Claudio Plaisant, Eulalie Lasseaux, Vincent Michaud, Isabelle Drumare, Benoit Arveiler","doi":"10.1016/j.gene.2024.149079","DOIUrl":"https://doi.org/10.1016/j.gene.2024.149079","url":null,"abstract":"<p><p>Oculocutaneous albinism type 1 is caused by variants in the TYR (tyrosinase) gene. We describe a family with two affected sibs who inherited the pathogenic missense TYR variant c.1146C > A;p.(Asn382Lys) from their mother and a deletion encompassing 65 kilobase pairs of the upstream region of the gene between hg38 coordinates chr11:89110944 and chr11:89175770, from their father. The deletion likely arose by non-homologous recombination since the regions including the two deletion breakpoints share no sequence homology. The deletion contains a single enhancer element that is homologous to a 5' Tyr core regulatory element in the mouse. A luciferase reporter assay showed that this element had a positive regulatory activity. This represents to our knowledge the first deletion solely restricted to non-coding upstream sequences of the TYR gene. It is assumed that the deletion down-regulates expression of the TYR gene and is therefore pathogenic, allowing to establish the diagnosis of OCA 1 in the patients. This study underscores the need to extend the search for pathogenic variants to regulatory regions either by whole genome sequencing or by targeted next generation sequencing of a panel including entire genes (exons, introns, flanking sequences) in order to improve the diagnostic rate in patients with albinism.</p>","PeriodicalId":12499,"journal":{"name":"Gene","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142603965","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
GenePub Date : 2024-11-04DOI: 10.1016/j.gene.2024.149076
Quanlin Mei, Ming Li, Jing Chen, Jiaxin Yang, Dingyue Duan, Jie Yang, Fengwang Ma, Ke Mao
{"title":"Genome-wide analyses of Ariadne family genes reveal their involvement in abiotic stress responses in apple.","authors":"Quanlin Mei, Ming Li, Jing Chen, Jiaxin Yang, Dingyue Duan, Jie Yang, Fengwang Ma, Ke Mao","doi":"10.1016/j.gene.2024.149076","DOIUrl":"10.1016/j.gene.2024.149076","url":null,"abstract":"<p><p>E3 ligases are essential for ubiquitination and play a role in regulating various aspects of eukaryotic life. Ariadne (ARI) proteins, a subfamily of RBR (RING-between-RING) proteins, have been recognized as a new class of RING-finger E3 ligases. Recent research has shed light on their potential involvement in plants' responses to abiotic stress. However, comprehensive studies on ARI genes in apple (Malus domestica) are still lacking. This study identified ten MdARI genes in the apple genome, and examined intraspecific and interspecific collinearity to explore the evolutionary relationships of ARI family members. Phylogenetic analyses classified MdARIs into two subfamilies (A and B), and by integrating gene structure, conserved motifs, and sequence comparison results, subfamily B was further divided into two subgroups (I and II). Tissue expression analyses revealed varied expression patterns of MdARI genes in different tissues, and subcellular localization showed that MdARI1-1, MdARI1-2, and MdARI9-1 were located in the nucleus, while the other seven MdARIs were distributed throughout the cell. Analyses of promoter cis-elements and expression patterns under cold, salt, and drought treatments indicated the involvement of MdARIs in abiotic stress responses. Several proteins crucial to the plant stress response were predicted to be potential MdARIs-interacting proteins based on the protein interaction network. Additionally, the interaction between UBC11 (E2) and MdARI7-2 was confirmed by a yeast two-hybrid (Y2H) experiment, suggesting that MdARI7-2 may function as an E3. These findings will greatly benefit future research on the role and mechanisms of ARI proteins in apple stress response.</p>","PeriodicalId":12499,"journal":{"name":"Gene","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142590539","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Identification of oxidative stress-related hub genes for predicting prognosis in diffuse large B-cell lymphoma.","authors":"Kewei Zhao, Qiuyue Wen, Qiuhui Li, Pengye Li, Tao Liu, Fang Zhu, Qiaoyun Tan, Liling Zhang","doi":"10.1016/j.gene.2024.149077","DOIUrl":"10.1016/j.gene.2024.149077","url":null,"abstract":"<p><strong>Background: </strong>Oxidative stress is a cellular characteristic that might induce the proliferation and differentiation of tumor cells and promote tumor progression in diffuse large B-cell lymphoma (DLBCL).</p><p><strong>Methods: </strong>The DLBCL gene sequencing dataset, tumor mutation burden data, copy number variation data of Somatic cell mutation data in TCGA were downloaded for data training analysis, along with four DLBCL datasets in GEO for validation analysis. The known oxidative stress related genes (OSRGs) were collected from websites. The weighted gene co-expression network analysis (WGCNA) was conducted on the TCGA DLBCL dataset to obtain gene modules related to oxidative stress and intersected with the known OSRGs to obtain the hub genes, which were used to perform consensus clustering on the samples to obtain new phenotypes. Next, the prognosis related OSRGs were selected through regression analysis algorithms and key genes were identified. These genes were used to establish the prognostic risk model and predictive model, and to compare functional and pathway differences among different risk groups.</p><p><strong>Results: </strong>Through website search, we obtained 297 known OSRGs, and after intersecting with WGCNA results, we obtained 26 OSRGs. The TCGA-DLBC samples were clustered into 2 subtypes with these genes and there were significant differences in immune infiltration between subtypes. After regression analysis, we obtained a total of four key genes, BMI1, CDKN1A, NOX1, and SESN1. The risk prediction model established with these four genes as variables has accurate prognostic prediction ability. The key genes interact with 65 miRNAs, 57 TFs, 47 RBPs, and 62 drugs, respectively, and are closely related to immune infiltration of the disease. Among them, CDKN1A and SESN1 had the highest variability.</p><p><strong>Conclusions: </strong>The key genes involved in oxidative stress could predict the prognosis of DLBCL and potentially become therapeutic targets.</p>","PeriodicalId":12499,"journal":{"name":"Gene","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142582509","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
GenePub Date : 2024-11-03DOI: 10.1016/j.gene.2024.149032
Can Mou, Lan Zhou, Jiao Jiao Xiong, Ling Lei
{"title":"A unique case of neurodevelopmental disorders and epilepsy linked to WDR45 variant inheritance and maternal mosaicism.","authors":"Can Mou, Lan Zhou, Jiao Jiao Xiong, Ling Lei","doi":"10.1016/j.gene.2024.149032","DOIUrl":"https://doi.org/10.1016/j.gene.2024.149032","url":null,"abstract":"<p><p>This paper reports a case of a WDR45 variant inherited from an asymptomatic low-percentage mosaic mother. The proband boy was found to have significant psychomotor developmental delay, epilepsy, and abnormal liver function at four months of age, and a hemizygous variant WDR45 c.867_869dupGTA (p.Y290*) was detected by high throughput sequencing, which has an ACMG rating of likely pathogenic variant. The same variant was detected by high-throughput sequencing of the amniotic fluid of the fetus at his mother's next pregnancy. Eventually, the same variant was detected in mosaic status in the unaffected mother by target capture-based deep sequencing of the asymptomatic mother, with a mutation load of 4.06 %.</p>","PeriodicalId":12499,"journal":{"name":"Gene","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142582421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"PICH, A protein that maintains genomic stability, can promote tumor growth.","authors":"Zeyuan Li, Wentao Zhang, Zihan Zhang, Guoming Mao, Linping Qi, Yubin Wang, Hanteng Yang, Huili Ye","doi":"10.1016/j.gene.2024.149074","DOIUrl":"10.1016/j.gene.2024.149074","url":null,"abstract":"<p><p>Genomic instability is regardedas a hallmark of cancer cells. It can be presented in many ways, among which chromosome instability has received attention. Ultrafine anaphase bridges are a typeof chromatin bridges, the untimely resolution of which can also lead to chromosome instability. PICH can play a role in maintaining chromosome stability by regulating chromosome morphologyand resolving ultrafine anaphase bridges. Recently, PICH has been found to be overexpressed in various cancers. Overexpression of PICH is related to the proliferation of tumors and poor prognosis. In this article, we consider that PICH can maintain genomic stability by regulating appropriate chromosome structure, ensuring proper chromosome segregation, and facilitating replication fork reversal. We summarize how PICH regulates chromosome stability, how PICH resolves Ultrafine anaphase bridges with other proteins, and how PICH promotes tumor progression.</p>","PeriodicalId":12499,"journal":{"name":"Gene","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142568130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
GenePub Date : 2024-11-02DOI: 10.1016/j.gene.2024.149066
Osama H. Abusara , Alaa M. Hammad , Rasha Debas , Eveen Al-Shalabi , Mohammed Waleed , F. Scott Hall
{"title":"The inflammation and oxidative status of rat lung tissue following smoke/vapor exposure via E-cigarette, cigarette, and waterpipe","authors":"Osama H. Abusara , Alaa M. Hammad , Rasha Debas , Eveen Al-Shalabi , Mohammed Waleed , F. Scott Hall","doi":"10.1016/j.gene.2024.149066","DOIUrl":"10.1016/j.gene.2024.149066","url":null,"abstract":"<div><h3>Background</h3><div>Tobacco smoking is a major worldwide health issue that contributes to millions of deaths annually. Electronic cigarettes (E-cigarettes) are also harmful. Smoke/vapor from E-cigarettes and tobacco products consists of free radicals and other toxic substances. Tissue damage in smokers, such as lungs, is highly observed and is linked to oxidative damage and inflammation.</div></div><div><h3>Methods</h3><div>The inflammation and oxidative status of rat lung tissues was examined following whole-body smoke/vapor exposure via E-cigarette, cigarette, and waterpipe for 2 h daily, 5 days per week for 8 weeks.</div></div><div><h3>Results</h3><div>Lung tissue damage was higher in cigarettes and waterpipe groups compared to the E-cigarette group. Collectively, there was a significant increase (p < 0.05) in the mRNA expression of pro-inflammatory mediators (<em>TNF-α</em>, <em>NF-κB</em>, <em>IL-1β</em>) with the exception of <em>IL-1β</em> in the E-cigarettes group. As for the anti-inflammatory mediators (<em>Nrf2</em> and <em>IL-10</em>), a significant reduction (p < 0.05) of mRNA expression was observed with the exception of <em>Nrf2</em> in the E-cigarette group. As for <em>IL-6</em>, there was a significant increase in its mRNA expression (p < 0.05) in the cigarette and waterpipe groups. There was also a significant decrease (p < 0.05) in the antioxidant activity of all antioxidants tested (GPx, SOD, and CAT) in all groups with the exception of SOD in the cigarette group.</div></div><div><h3>Conclusion</h3><div>Smoke/vapor administered via E-cigarette, cigarette, and waterpipe elicits inflammation and oxidative stress in rat lungs that is accompanied by histopathological changes.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142568134","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"CRISPR applications in microbial World: Assessing the opportunities and challenges","authors":"Farhan Kursheed , Esha Naz , Sana Mateen , Ume Kulsoom","doi":"10.1016/j.gene.2024.149075","DOIUrl":"10.1016/j.gene.2024.149075","url":null,"abstract":"<div><div>Genome editing has emerged during the past few decades in the scientific research area to manipulate genetic composition, obtain desired traits, and deal with biological challenges by exploring genetic traits and their sequences at a level of precision. The discovery of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) as a genome editing tool has offered a much better understanding of cellular and molecular mechanisms. This technology emerges as one of the most promising candidates for genome editing, offering several advantages over other techniques such as high accuracy and specificity. In the microbial world, CRISPR/Cas technology enables researchers to manipulate the genetic makeup of micro-organisms, allowing them to achieve almost impossible tasks. This technology initially discovered as a bacterial defense mechanism, is now being used for gene cutting and editing to explore more of its dimensions. CRISPR/Cas 9 systems are highly efficient and flexible, leading to its widespread uses in microbial research areas. Although this technology is widely used in the scientific community, many challenges, including off-target activity, low efficiency of Homology Directed Repair (HDR), and ethical considerations, still need to be overcome before it can be widely used. As CRISPR/Cas technology has revolutionized the field of microbiology, this review article aimed to present a comprehensive overview highlighting a brief history, basic mechanisms, and its application in the microbial world along with accessing the opportunities and challenges.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142568068","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Integrating multiomics sequencing analyses uncover the key mechanisms related to oxidative stress, mitochondria, and immune cells in keloid.","authors":"Lianbo Zhang, Ruizhu Liu, Mingxi Li, Guang Zhang, Zichao Wang, Haiyan Qin","doi":"10.1016/j.gene.2024.149078","DOIUrl":"10.1016/j.gene.2024.149078","url":null,"abstract":"<p><strong>Background: </strong>This study aimed to investigate the key molecular mechanisms underlying keloid pathogenesis by integrating oxidative stress, mitochondria, and immune cells.</p><p><strong>Methods: </strong>Transcriptome sequencing (mRNA, lncRNA, and circRNA expression data), proteomic sequencing, and small RNA sequencing analyses of lesional and non-lesional skin of patients with keloids and healthy control (normal) skin were conducted. By integrating mRNA and publicly available gene expression data (GSE158395), differentially expressed genes related to oxidative stress and mitochondrial function in keloids were identified. Hub genes were identified using various bioinformatics analyses such as immune infiltration analysis, weighted gene co-expression network analysis, machine learning, and expression validation using proteomics sequencing data. Moreover, a competing endogenous RNA (ceRNA) network of hub genes was constructed by combining miRNA, lncRNA, and circRNA expression data. Five hub genes were identified: MGST1, DHCR24, ALDH3A2, ADH1B, and FKBP5.</p><p><strong>Results: </strong>These hub genes had a high diagnostic value for keloids, with an AUC value > 0.8 each. In addition, five hub genes were associated with the infiltration of multiple immune cells. The immune cells with the strongest positive and negative correlations with hub genes were M0 and M1 macrophages. A ceRNA network was constructed, and several ceRNAs, such as AC005062.1/miR-134-5p/FKBP5 and BASP1-AS1/miR-503-5p/ADH1B, were identified. These five hub genes may contribute to keloid pathogenesis.</p><p><strong>Conclusion: </strong>These genes and their related ceRNAs may serve as diagnostic biomarkers and therapeutic targets for keloids.</p>","PeriodicalId":12499,"journal":{"name":"Gene","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142568111","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
GenePub Date : 2024-11-01DOI: 10.1016/j.gene.2024.149073
Sedef Erkunt Alak , Ceren Gül , Mervenur Güvendi , Aytül Gül , Muhammet Karakavuk , Aysu Değirmenci Döşkaya , Seren Kaplan , Cemal Ün , Adnan Yüksel Gürüz , Mert Döşkaya , Hüseyin Can
{"title":"An optimized ROP6 mRNA construct successfully expressed immunogenic Toxoplasma gondii ROP6 protein in cell culture","authors":"Sedef Erkunt Alak , Ceren Gül , Mervenur Güvendi , Aytül Gül , Muhammet Karakavuk , Aysu Değirmenci Döşkaya , Seren Kaplan , Cemal Ün , Adnan Yüksel Gürüz , Mert Döşkaya , Hüseyin Can","doi":"10.1016/j.gene.2024.149073","DOIUrl":"10.1016/j.gene.2024.149073","url":null,"abstract":"<div><div><em>Toxoplasma gondii</em> is an apicomplexan parasite infecting all mammals including humans and causes toxoplasmosis. There is no vaccine available for humans and thus vaccine development efforts continue using novel antigens and/or vaccine platforms. Since our previous microarray screening study showed that ROP6 is a suitable antigen to be used in vaccine studies, in this study, we aimed to design an optimized mRNA construct encoding the ROP6 protein and then demonstrate its efficiency and immunogenicity using <em>in vitro</em> methods. For this, we constructed a pT7CFE1-Chis/ROP6 vector encoding optimized ROP6 mRNA containing EMCV 5′UTR with IRES and a 20 nucleotides fragment from alpha globin 3′ UTR. Then, we generated the optimized ROP6 mRNAs with anti-reverse cap analogue (ARCA) and approximately 150 nucleotide long poly-A tail. Next, HEK293T cells were transfected with the optimized ROP6 mRNAs to show recombinant ROP6 protein expression capability. Moreover, we expressed <em>in vitro</em> recombinant ROP6 protein in HeLa cell lysate using the pT7CFE1-Chis/ROP6 vector to reveal the immunogenicity of recombinant ROP6 protein using sera samples collected from mice infected with PRU strain of <em>T. gondii</em>. The IFA and Western blot results showed that the optimized ROP6 mRNAs successfully expressed the recombinant ROP6 protein in HEK293T cells. Moreover the recombinant ROP6 protein expressed in HeLa cell lysate strongly reacted with sera samples collected from mice<em>.</em> The absorbance difference detected among positive and negative mice serum samples analyzed was statistically significant, indicating that the recombinant ROP6 protein was immunogenic (<em>P</em> = 0.0003). In conclusion, this study demonstrated that the optimized ROP6 mRNAs can be used in the development of mRNA vaccines against toxoplasmosis.</div></div>","PeriodicalId":12499,"journal":{"name":"Gene","volume":null,"pages":null},"PeriodicalIF":2.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142568058","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}