Molekulyarnaya Biologiya最新文献

[Dynamics of Transgene Expression in the MVA Vaccine Vector under the Control of the p11, p13.5, pLEO160, p7.5, and mH5 Promoters and Independence of the Transgene Expression Level from the Insertion Locus]. [p11、p13.5、pLEO160、p7.5和mH5启动子控制下MVA疫苗载体中转基因表达的动态及转基因表达水平与插入位点的独立性]。

Molekulyarnaya Biologiya Pub Date : 2025-03-01 DOI: 10.31857/S0026898425020065, EDN: GGEKOK

O V Orlova, D V Glazkova, O N Sidorova, F A Urusov, G A Shipulin, E V Bogoslovskaya

{"title":"[Dynamics of Transgene Expression in the MVA Vaccine Vector under the Control of the p11, p13.5, pLEO160, p7.5, and mH5 Promoters and Independence of the Transgene Expression Level from the Insertion Locus].","authors":"O V Orlova, D V Glazkova, O N Sidorova, F A Urusov, G A Shipulin, E V Bogoslovskaya","doi":"10.31857/S0026898425020065, EDN: GGEKOK","DOIUrl":"https://doi.org/10.31857/S0026898425020065, EDN: GGEKOK","url":null,"abstract":"The modified vaccinia virus Ankara (MVA), characterized by high immunogenicity and proven safety, is considered a promising vector for vaccine development. An undeniable advantage of the MVA vector is its high capacity and the ability to incorporate several transgenes into different loci of the viral genome, enabling the creation of multivalent vaccines that encode multiple antigens simultaneously. This study examined the expression of a transgene encoding influenza virus protein epitopes after its integration into five loci of the MVA genome. The results demonstrated that the level of transgene expression is independent of the integration locus. Additionally, the dynamics of the expression of a reporter gene encoding the Enhanced Green Fluorescent Protein (EGFP) were determined under the control of the vaccinia virus promoters p11, p13.5, pLEO160, p7.5, and mH5 upon insertion of the expression cassette into the F13L gene locus of MVA. The highest expression level, though with a delayed onset of protein synthesis, was achieved with the late promoter p11. Using the p13.5 promoter resulted in earlier synthesis of EGFP in cells and higher gene expression level compared to the promoters pLEO160, p7.5, and mH5, which provided similar levels and dynamics of the reporter gene expression. These findings may be useful for developing multi-antigenic MVA-vectored vaccines.","PeriodicalId":39818,"journal":{"name":"Molekulyarnaya Biologiya","volume":"59 2","pages":"244-254"},"PeriodicalIF":0.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144486432","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[MLE (DHX9) Helicase Regulates the Expression of Constitutive and Inducible Isoforms of the Conserved Nuclear Receptor FTZ-F1 (NR5A3)]. [MLE （DHX9）解旋酶调控保守核受体FTZ-F1 （NR5A3）的组成型和诱导型异构体的表达]。

Molekulyarnaya Biologiya Pub Date : 2025-03-01 DOI: 10.31857/S0026898425020089, EDN: GFZUGJ

J V Nikolenko, S G Georgieva

{"title":"[MLE (DHX9) Helicase Regulates the Expression of Constitutive and Inducible Isoforms of the Conserved Nuclear Receptor FTZ-F1 (NR5A3)].","authors":"J V Nikolenko, S G Georgieva","doi":"10.31857/S0026898425020089, EDN: GFZUGJ","DOIUrl":"https://doi.org/10.31857/S0026898425020089, EDN: GFZUGJ","url":null,"abstract":"In addition to participating in dosage compensation, the MLE helicase in D. melanogaster performs many functions in the regulation of gene expression, as does its human ortholog DHX9. Many of these functions are evolutionarily conserved and poorly explored. MLE has previously been shown to be involved in the regulation of inducible transcription of the ftz-f1 gene encoding the evolutionarily conserved nuclear receptor NR5A3. The ftz-f1 gene also encodes a constitutive transcript synthesized from an alternative promoter. The present work is devoted to the investigation of the role of MLE in the regulation of constitutive transcription of the ftz-f1 gene. This work shows that in S2 cell culture, MLE binds to the constitutive promoter and controls both inducible and constitutive transcription of the ftz-f1 gene. A novel MLE-binding cis- regulatory element of the ftz-f1 gene, enhancer 663, was identified. Using chromosome conformation capture technique the interaction of enhancer 663 with constitutive and inducible promoters of ftz-f1 gene in S2 cell culture was demonstrated. Examination of enhancer 663 histone H3 acetylation showed that it is involved in the activity of both promoters. Knockdown of MLE in S2 cell culture causes an increase in constitutive transcription. The effect of MLE on transcription beyond dosage compensation in vivo at the adult stage was shown for the first time. It was shown that at the adult stage MLE binds to both inducible and constitutive promoters and to enhancer 663. Mutation in the mle gene leads to increased expression of both transcripts of the ftz-f1 gene in females. The data obtained are important for understanding and further study of the evolutionarily conserved functions of MLE and its human ortholog DHX9.","PeriodicalId":39818,"journal":{"name":"Molekulyarnaya Biologiya","volume":"59 2","pages":"266-276"},"PeriodicalIF":0.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144486445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Optimization of Cytotoxic Properties of Magnetic Nanoparticle- Based Doxorubicin Delivery System]. 磁性纳米颗粒基阿霉素给药系统细胞毒性的优化研究。

Molekulyarnaya Biologiya Pub Date : 2025-03-01 DOI: 10.31857/S0026898425020108, EDN: GFXGJH

A I Kurtova, A V Svetlakova, O A Kolesnikova, V O Shipunova

{"title":"[Optimization of Cytotoxic Properties of Magnetic Nanoparticle- Based Doxorubicin Delivery System].","authors":"A I Kurtova, A V Svetlakova, O A Kolesnikova, V O Shipunova","doi":"10.31857/S0026898425020108, EDN: GFXGJH","DOIUrl":"https://doi.org/10.31857/S0026898425020108, EDN: GFXGJH","url":null,"abstract":"Doxorubicin (DOX) is a widely used cytotoxic drug known for its high antitumor activity; however, its use is associated with side effects. The development of DOX delivery systems that can minimize systemic toxicity and enhance therapeutic efficacy is an urgent task in modern oncology. We studied the process of loading nanoparticles (NPs) with DOX under conditions that promote DOX precipitation to achieve maximum sorption efficiency. For this purpose, polymer-stabilized magnetic NPs were synthesized, and the efficiency of loading and sedimentation was examined based on the buffer type, DOX concentration, and incubation time with the drug. Our findings indicated that in solutions with the most pronounced DOX sedimentation (phosphate and borate buffers), loading was most effective. In a phosphate buffer with an initial DOX concentration of 667 μg/mL, the loading was 886 mg DOX/g NP. The sorption of DOX on NPs under these conditions reached 85% within the first hour and increased to 90% within 3 hours. The release of DOX from NPs was 25% at pH 7.4 and 96% at pH 5.4. Analysis of the survival of EMT-HER2 breast cancer cells demonstrated that the cytotoxicity of NPs loaded with DOX under sedimentation conditions was eight times higher than that of NPs loaded at a concentration of 20 μg/mL, where DOX did not form a sediment. These results suggest that NPs loaded with DOX under sedimentation conditions can be considered an effective delivery system that not only maintains the cytotoxic properties of DOX but also significantly enhances the content and release of the delivered drug.","PeriodicalId":39818,"journal":{"name":"Molekulyarnaya Biologiya","volume":"59 2","pages":"288-298"},"PeriodicalIF":0.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144486446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Improving the Efficiency and Safety of Human CCR5 Gene Editing by Selection of Optimal Guide RNAs for SpCAS9 and CAS12A]. [通过选择SpCAS9和CAS12A的最佳引导rna提高人类CCR5基因编辑的效率和安全性]。

Molekulyarnaya Biologiya Pub Date : 2025-03-01 DOI: 10.31857/S0026898425020055, EDN: GGKOBN

R R Mintaev, D V Glazkova, J A Taran, E V Bogoslovskaya, G A Shipulin

引用次数: 0

[Sample Preparation and Sequencing Efficiency of microRNA Libraries from Pituitary Adenoma Tissue and Blood Plasma of Patients with Acromegaly for the Illumina Platform]. [用于Illumina平台的垂体腺瘤组织和肢端肥大症患者血浆microRNA文库的样品制备和测序效率]。

Molekulyarnaya Biologiya Pub Date : 2025-03-01 DOI: 10.31857/S0026898425020121, EDN: GFUSPR

E V Ignatieva, E S Nerubenko, O A Ivanova, U A Tsoy, R I Dmitrieva

{"title":"[Sample Preparation and Sequencing Efficiency of microRNA Libraries from Pituitary Adenoma Tissue and Blood Plasma of Patients with Acromegaly for the Illumina Platform].","authors":"E V Ignatieva, E S Nerubenko, O A Ivanova, U A Tsoy, R I Dmitrieva","doi":"10.31857/S0026898425020121, EDN: GFUSPR","DOIUrl":"https://doi.org/10.31857/S0026898425020121, EDN: GFUSPR","url":null,"abstract":"MicroRNAs in tissues and biological fluids represent a promising class of biomarkers for the molecular diagnostics and therapy of numerous diseases, including oncological diseases. Biomarkers based on easily accessible biological fluids, primarily blood-based biomarkers, are of particular value for diagnostic and prognostic purposes. To explore the potential of microRNAs as prognostic cancer markers and targets for molecular therapy, global microRNA profiling is required, which is provided by next-generation sequencing (NGS). NGS offers high sensitivity, single nucleotide resolution, and the possibility of profiling a large number of samples in parallel. Despite the promising potential of microRNAs as biomarkers and the growing number of works in this area, the literature does not address in sufficient detail the problems associated with sample preparation methods, the specifics of library preparation for microRNA sequencing, and the difficulties of quantitative analysis. Protocols for creating libraries for microRNA sequencing present specific challenges and require selecting conditions for each type of biological sample. Here, we present in detail the preparation of libraries for microRNA sequencing from pituitary adenoma tumor tissue and blood plasma of patients with acromegaly on the Illumina platform. We discuss the difficulties and limitations of the methods and evaluate the effectiveness of sequencing plasma and brain samples. This work can serve as a guide for researchers studying the mechanisms of microRNA regulation in endocrine diseases of the pituitary gland and will also allow for the adaptation of technical procedures for various biological samples in relation to other pathologies.","PeriodicalId":39818,"journal":{"name":"Molekulyarnaya Biologiya","volume":"59 2","pages":"309-323"},"PeriodicalIF":0.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144486449","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Identification of Genetic Markers of Predisposition to Thrombogenic Diseases by Minisequencing Analysis: Reagent Set "SNP2-TMG"]. [通过微序列分析鉴定血栓性疾病易感性的遗传标记：试剂盒“SNP2-TMG”]。

Molekulyarnaya Biologiya Pub Date : 2025-03-01 DOI: 10.31857/S0026898425020031, EDN: GGTFSD

A V Grudo, I V Haidukevich, G V Sergeev

{"title":"[Identification of Genetic Markers of Predisposition to Thrombogenic Diseases by Minisequencing Analysis: Reagent Set \"SNP2-TMG\"].","authors":"A V Grudo, I V Haidukevich, G V Sergeev","doi":"10.31857/S0026898425020031, EDN: GGTFSD","DOIUrl":"https://doi.org/10.31857/S0026898425020031, EDN: GGTFSD","url":null,"abstract":"Thrombogenic risk factors (blood coagulation disorders and thrombophilia) are the cause of cardiovascular diseases, among which genetic factors are worth highlighting (genetic polymorphism of the blood coagulation system, angiogenesis factors, and components of the lipid metabolism system). Early identification of clinically significant polymorphisms in genes that cause predisposition to thrombogenic diseases allows for preventive measures and timely diagnosis even before the onset of the clinical picture of the disease, and for patients with an already confirmed diagnosis, genetic diagnostics makes it possible to check the hereditary nature of the disease, select treatment tactics, and predict the risk of developing of adverse drug reactions. This article describes the process of developing the \"SNP2-TMG\" kit, designed to identify ten genetic markers of susceptibility to thrombogenic diseases (rs1801131, rs6025, rs11549465, rs429358, rs7412, rs1799963, rs6050, rs1799762, rs2010963, and rs1801133), by the minisequencing technique (SNaPshot technology). This kit has passed clinical trials and is approved for medical use.","PeriodicalId":39818,"journal":{"name":"Molekulyarnaya Biologiya","volume":"59 2","pages":"201-211"},"PeriodicalIF":0.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144486434","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Transcription Factor PAX4: Role in Differentiation of Insulin- Producing β Cells during Pancreas Development and Association with Diabetes]. [转录因子PAX4：胰腺发育过程中胰岛素生成β细胞分化的作用及与糖尿病的关系]。

Molekulyarnaya Biologiya Pub Date : 2025-03-01 DOI: 10.31857/S0026898425020023, EDN: GGXQEP

A I Melnikova, T S Krasnova, P M Rubtsov

{"title":"[Transcription Factor PAX4: Role in Differentiation of Insulin- Producing β Cells during Pancreas Development and Association with Diabetes].","authors":"A I Melnikova, T S Krasnova, P M Rubtsov","doi":"10.31857/S0026898425020023, EDN: GGXQEP","DOIUrl":"https://doi.org/10.31857/S0026898425020023, EDN: GGXQEP","url":null,"abstract":"PAX4 (Paired Box 4) is a transcription factor that is expressed mainly in the pancreas and plays a key role in the development of insulin-producing β cells at the embryonic stage. In mature cells, PAX4 acts as a main regulator of their adaptation under pathological conditions. The importance of PAX4 to the proper function of pancreatic islets has been demonstrated in studies of the relationship between mutations of the PAX4 gene and various forms of diabetes mellitus (DM). PAX4 overexpression in adult islets stimulates β-cell proliferation and resistance to apoptosis. Taken together, the data indicate that PAX4 provides a potential target to develop new DM treatments aimed at reprogramming different cell types into insulin-producing cells and promoting their proliferation to replenish the β-cell pool lost during disease progression. The development of such methods requires knowledge of the molecular mechanisms that control expression of PAX4 and its target genes. The review summarizes the data on the structure and expression of the human PAX4 gene. Interactions of various transcription factors during differentiation of pancreatic cells and the formation of islets of Langerhans are described along with the role of PAX4 in the processes. Associations between mutations of human PAX4 and various DM forms were considered. A final part of the review examines the prospects for reprogramming cells of other types into insulin-producing cells and discusses the effects on PAX4- regulated signaling pathways as a means to develop new approaches to DM treatment.","PeriodicalId":39818,"journal":{"name":"Molekulyarnaya Biologiya","volume":"59 2","pages":"189-200"},"PeriodicalIF":0.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144486450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[πDMD Simulation as a Strategy for Refinement of AlphaFold2 Modeled Fuzzy Protein Complexes Structures]. [πDMD模拟作为AlphaFold2模型模糊蛋白复合物结构的改进策略]。

Molekulyarnaya Biologiya Pub Date : 2025-03-01 DOI: 10.31857/S0026898425020095, EDN: GFYXKD

N G Muradyan, A A Sargsyan, V G Arakelov, A K Paronyan, G G Arakelov, K B Nazaryan

引用次数: 0

[Efficient Method to Isolate High-Quality RNA from Mycelia of Toxigenic Fungi Fusarium sp.] 从产毒真菌Fusarium sp.菌丝体中分离高质量RNA的高效方法

Molekulyarnaya Biologiya Pub Date : 2025-03-01 DOI: 10.31857/S0026898425020138, EDN: GFUSAT

A A Stakheev, D Yu Ryazantsev, N G Gabrielyan, A V Poluboyarinova, M E Taliansky, S K Zavriev

引用次数: 0

[Influence of Homology Arm Length and Structure on the Efficiency of Long Transgene Integration into a Cleavage Site Induced by SpCas9 or AsCpf1]. [同源臂长和结构对SpCas9或AsCpf1诱导的长转基因整合到切割位点效率的影响]。

Molekulyarnaya Biologiya Pub Date : 2025-03-01 DOI: 10.31857/S0026898425020079, EDN: GGBDOW

J A Taran, R R Mintaev, D V Glazkova, B V Belugin, E V Bogoslovskaya, G A Shipulin

{"title":"[Influence of Homology Arm Length and Structure on the Efficiency of Long Transgene Integration into a Cleavage Site Induced by SpCas9 or AsCpf1].","authors":"J A Taran, R R Mintaev, D V Glazkova, B V Belugin, E V Bogoslovskaya, G A Shipulin","doi":"10.31857/S0026898425020079, EDN: GGBDOW","DOIUrl":"https://doi.org/10.31857/S0026898425020079, EDN: GGBDOW","url":null,"abstract":"One of the promising new approaches to the treatment of HIV infection is CRISPR/Cas-mediated knockout of the CCR5 receptor gene followed by the integration of an anti-HIV gene into the break site. Numerous studies have focused on the knockout of the CCR5 gene; however, the efficiency of subsequent targeted integration of long fragments remains poorly studied. To evaluate the efficiency of this approach, we used HT1080 cells and investigated the integration of a cassette expressing the EGFP gene into the CCR5 locus using two different nucleases (SpCas9 and AsCpf1) and various donor DNA constructs delivered by recombinant adeno-associated viral vectors (rAAV). For each nuclease, we designed five variants of donor DNA differing in the length (ranging from 150 to 1000 bp) or structure of the homology arms. The efficiency of transgene integration with 150 bp homology arms was the lowest for both nucleases and differed significantly from constructs with longer homology arms. Furthermore, it was shown that the presence of nuclease cleavage sites in the donor DNA flanking the cassette with homology arms did not affect the efficiency of transgene integration during AAV delivery. We demonstrated that the AsCpf1 nuclease provided higher efficiency of EGFP transgene integration than SpCas9, despite the lower efficiency of CCR5 knockout. The maximum percentage of cells with the integrated transgene was achieved using the AsCpf1 nuclease and an expression cassette with 600 bp homology arms, reaching 59 ± 6%.","PeriodicalId":39818,"journal":{"name":"Molekulyarnaya Biologiya","volume":"59 2","pages":"255-265"},"PeriodicalIF":0.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144486436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0