Gene ReportsPub Date : 2024-11-01DOI: 10.1016/j.genrep.2024.102079
Joel Henrique Ellwanger, José Artur Bogo Chies
{"title":"Toxicogenomics supports carcinogenic action of tattoo ink components","authors":"Joel Henrique Ellwanger, José Artur Bogo Chies","doi":"10.1016/j.genrep.2024.102079","DOIUrl":"10.1016/j.genrep.2024.102079","url":null,"abstract":"<div><div>Growing evidence suggests that tattoos may be associated with an increased risk of cancer development due to carcinogenic components present in tattoo inks. We explored this issue using The Comparative Toxicogenomics Database. Exploratory toxicogenomic data corroborate the association between cancer and tattoo ink components, especially concerning the effects of polycyclic aromatic hydrocarbons (PAH). The top-15 genes affected by PAH and the top-15 diseases associated with PAH were listed. Polycyclic aromatic hydrocarbons and other components present in tattoo inks affect the expression of multiple genes that participate in the metabolism of xenobiotics, cell death, and immune responses, and disruption of these processes may facilitate carcinogenesis. In agreement, cancer is the main disease category associated with PAH. In Brazil and other countries, there are significant deficiencies in the regulation, marketing, and inspection of substances used in tattoo inks. Considering the immense number of individuals who get tattoos around the world, tattoo inks should be subjected to more complete toxicological studies, and stricter regulation of tattoo ink usage is needed.</div></div>","PeriodicalId":12673,"journal":{"name":"Gene Reports","volume":null,"pages":null},"PeriodicalIF":1.0,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142587268","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}
{"title":"Comparative RNA-seq analysis reveals transposable element-mediated transcriptional reprogramming under phosphorus-starvation stress in rice (Oryza sativa L.)","authors":"Simardeep Kaur , Karishma Seem , K.K. Vinod , Dwijesh Chandra Mishra , Suresh Kumar , Trilochan Mohapatra","doi":"10.1016/j.genrep.2024.102077","DOIUrl":"10.1016/j.genrep.2024.102077","url":null,"abstract":"<div><div>Phosphorus (P) deficiency hinders crop productivity of 50 % of the rice grown in Asia, Africa, and South America. About 90 % of the phosphate in fertilizers applied to the crops gets fixed in the soil, reducing its availability to plants. This necessitates increased use of phosphatic fertilizers leading to higher cost of cultivation and environmental pollution. Although molecular mechanisms of P-deficiency tolerance in rice are being deciphered, the role of transposable elements (TEs) in transcriptional reprogramming under P-starvation/deficiency stress has not yet been reported. To investigate the role of <em>Pup1</em> QTL in controlling TE-mediated reprogramming of gene expression, a pair of contrasting rice [Pusa-44 and its Near-Isogenic Line (NIL)-23] genotypes were grown hydroponically under control and stressed (0 ppm Pi) conditions. Comparative RNA-seq analysis of root and shoot tissues from 45-day-old plants of the rice genotypes revealed TE-mediated transcriptional reprogramming affecting biological processes and cellular components. Significantly up-regulated expression of several TEs under P-starvation stress, controlled by <em>Pup1</em> QTL, particularly in shoots of NIL-23 indicates their crucial role in P homeostasis. Moreover, comparative physio-biochemical analyses confirmed the stress tolerance of NIL-23. Several biological processes including DNA replication/repair, metabolism, signaling, and phosphorylation were modulated through differential (mainly up-regulated) expression of TEs (controlled by <em>Pup1</em> QTL) in shoots of NIL-23 under P-starvation. To the best of our knowledge, this is a pioneer study on the role of TEs in reprogramming biological processes/molecular functions/cellular components involved in P-use efficiency in rice under stress. The findings advance our understanding of the functions of <em>Pup1</em> to improve the P-use efficiency/productivity of rice in P-deficient soils.</div></div>","PeriodicalId":12673,"journal":{"name":"Gene Reports","volume":null,"pages":null},"PeriodicalIF":1.0,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142573481","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}
Gene ReportsPub Date : 2024-10-31DOI: 10.1016/j.genrep.2024.102076
Rajender Goud Arolla , K. Srinivas Naik
{"title":"Genomic characterization and comparative genomics of Chlorella sp. CH2018 from Musi River water, India","authors":"Rajender Goud Arolla , K. Srinivas Naik","doi":"10.1016/j.genrep.2024.102076","DOIUrl":"10.1016/j.genrep.2024.102076","url":null,"abstract":"<div><div>Microalgae, a diverse class of photosynthetic eukaryote, can provide food and energy sustainably. Selecting productive strains is crucial for commercial viability. Here, we isolated an axenic microalgal species from Musi River water, and based on its morphology, molecular makeup, and cell wall composition identified it as <em>Chlorella</em> sp. CH2018. Scanning electron microscopy was used to measure the size of cells, which were found to be 3-, 4-, or 5-μm in diameter with a discernible thick outer cell wall. The characterization of algal genomes is imperative for comprehending species, studying metabolic pathways, and modifying genetics. In this study, we sequenced the entire genome of <em>Chlorella</em> sp. CH2018, yielded genome size of 56.83 Mb with 11,143 functionally annotated protein-coding gene models. The Gene Ontology (GO) analysis revealed that <em>Chlorella</em>'s predominant metabolic pathway is carbohydrate metabolism. Ortholog comparative analysis of species of phylum Chlorophyta with <em>Chlorella</em> sp. CH2018 showed that the isolated species possesses unique protein families with a maximum number of 6292 ortholog groups with <em>Chlorella sorokiniana</em>. The phylogenetic tree created by concatenating single-copy ortholog sequences demonstrates the uniqueness of <em>Chlorella</em> sp. CH2018, and its genome sequence serves as a genetic resource for future research.</div></div>","PeriodicalId":12673,"journal":{"name":"Gene Reports","volume":null,"pages":null},"PeriodicalIF":1.0,"publicationDate":"2024-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142587267","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}
Gene ReportsPub Date : 2024-10-30DOI: 10.1016/j.genrep.2024.102074
Hima J. Challa, Kalyan Ram Uppaluri, A.Sai. Rishika Gopikar, Rebecca Chalcedony, Srinivas Kethavath, K. Sri Manjari, K. Krishna Vardhani, Kalyani Palasamudram, Natya Kanuri, Aswini Korivepi
{"title":"A case report on genetic variants in CHD8, KMT2C, EP300, and TCF4 associated with autism spectrum disorder","authors":"Hima J. Challa, Kalyan Ram Uppaluri, A.Sai. Rishika Gopikar, Rebecca Chalcedony, Srinivas Kethavath, K. Sri Manjari, K. Krishna Vardhani, Kalyani Palasamudram, Natya Kanuri, Aswini Korivepi","doi":"10.1016/j.genrep.2024.102074","DOIUrl":"10.1016/j.genrep.2024.102074","url":null,"abstract":"<div><div>Autism spectrum disorder (ASD) is a complex neurodevelopmental condition influenced by several environmental and genetic factors. We present a case of a 7-year-old male with ASD, neurogenic voiding dysfunction (NVD), speech and developmental delays, and hyperactivity, and current treatment includes speech, occupational, and behavioral therapy. Family history includes consanguinity and maternal intellectual disability. Whole-exome sequencing (WES) identified maternally inherited variants in <em>CHD8</em> (benign), <em>KMT2C</em> (likely pathogenic), and <em>EP300 and TCF4</em> (uncertain significance). Despite the benign classification of the <em>CHD8</em> variant, its association with ASD highlights the complexity of genotype-phenotype correlations. The likely pathogenic <em>KMT2C</em> frameshift mutation and deletions in <em>EP300 and TCF4</em> suggest a multifactorial genetic basis for ASD in this patient. These findings highlight the importance of integrating clinical and genetic data for accurate diagnosis and personalized treatment. Whole Exome Sequencing (WES) analysis revealed these variants in the child's mother, uncle, and maternal grandfather, with the maternal uncle unaffected by ASD, ID, or ADHD, indicating potential variant interplay in disease manifestation. This case emphasizes the need for further research to elucidate the combined effects of these variants, enhancing our understanding of ASD's genetic landscape and improving clinical outcomes.</div></div>","PeriodicalId":12673,"journal":{"name":"Gene Reports","volume":null,"pages":null},"PeriodicalIF":1.0,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142573482","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}
Gene ReportsPub Date : 2024-10-30DOI: 10.1016/j.genrep.2024.102073
Aynaz Ghojoghi , Ali Zarei Mahmoudabadi , Sadegh Khodavaisy , Eisa Nazar , Mahnaz Fatahinia
{"title":"Unveiling the genotypic diversity of Candida albicans and Candida dubliniensis in the oral cavities of drug abusers in Ahvaz, Iran","authors":"Aynaz Ghojoghi , Ali Zarei Mahmoudabadi , Sadegh Khodavaisy , Eisa Nazar , Mahnaz Fatahinia","doi":"10.1016/j.genrep.2024.102073","DOIUrl":"10.1016/j.genrep.2024.102073","url":null,"abstract":"<div><div><em>Candida albicans</em> is a diploid yeast that, under certain conditions, can cause oral or oropharyngeal infections, particularly in immunocompromised individuals. Recent molecular investigations have classified genotypes A, B, and C for <em>Candida albicans</em>, along with genotype D for <em>Candida dubliniensis</em>. This study aimed to identify the different genotypes of the <em>C. albicans</em> complex in drug abusers in Iran. Oral swabs were collected from drug abusers and cultured on CHROMagar <em>Candida</em>. A 21-plex PCR method was employed for the detection of isolates, and the <em>Candida</em> 25S rDNA gene was amplified using primer pairs CA-INT-L and CA-INT-R for ABC genotyping of <em>C. albicans</em>. Out of the 245 substance abusers screened, 151 individuals (61.63 %) were found to harbor the <em>C. albicans</em> complex. The most common genotype among patients was genotype D (39.1 %), followed by genotype A (31.12 %), genotype B (9.93 %), and genotype C (5.29 %). Additionally, 14.56 % of patients had a mixed genotype. Notably, significant differences in genotype distribution were observed in relation to age, underlying diseases, and marital status (<em>P</em> < 0.05). This study highlights the significance of molecular genotyping in understanding the epidemiology of <em>C. albicans</em> and <em>C. dubliniensis</em> in at-risk groups.</div></div>","PeriodicalId":12673,"journal":{"name":"Gene Reports","volume":null,"pages":null},"PeriodicalIF":1.0,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142578312","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}
Gene ReportsPub Date : 2024-10-30DOI: 10.1016/j.genrep.2024.102075
Anqi Chen , Chaoneng Ji , Chengtao Li , Suhua Zhang
{"title":"Comparing liver RNA-seq analysis of human, rhesus monkey, mouse and rat","authors":"Anqi Chen , Chaoneng Ji , Chengtao Li , Suhua Zhang","doi":"10.1016/j.genrep.2024.102075","DOIUrl":"10.1016/j.genrep.2024.102075","url":null,"abstract":"<div><h3>Background</h3><div>Animal models are commonly utilized by the pharmaceutical industry to evaluate new drugs; however, the results of these studies are not always applicable to humans due to differences between the genetic backgrounds.</div></div><div><h3>Methods</h3><div>Since liver plays a prominent role in drug metabolism, we evaluated the expression profiles of human, mouse, rat, and rhesus monkey liver tissues. The differentially expressed genes were identified in different genders, strains, and species.</div></div><div><h3>Results</h3><div>The primates had fewer differences compared to the rodents, suggesting rhesus monkey could be a preferred model organism. Although the human HGFR (MET), a putative AAV3 receptor, together with its ligand HGF, showed no significant expression differences to the mammal models, the ligand-receptor (HGF/MET) ratio difference between the human (HGF/MET = 0.56) and the mice (HGF/MET = 0.12) might result in the transduction failure of murine hepatocytes using the AAV3 vectors. Our results also showed ACE2 expression was not detected in the human liver, while low expression levels were observed in other species, suggesting that the liver might not be the main target of SARS-CoV2.</div></div><div><h3>Conclusion</h3><div>In summary, even though all model organisms have limitations, non-human primates or humanized animals are indispensable during drug development before proceeding to clinical trials.</div></div>","PeriodicalId":12673,"journal":{"name":"Gene Reports","volume":null,"pages":null},"PeriodicalIF":1.0,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142573480","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}
Gene ReportsPub Date : 2024-10-29DOI: 10.1016/j.genrep.2024.102067
Sanjukta Dasgupta
{"title":"Identification and molecular modelling of potential drugs targeting the genes involved in the progression of lung cancer in patients with idiopathic pulmonary fibrosis","authors":"Sanjukta Dasgupta","doi":"10.1016/j.genrep.2024.102067","DOIUrl":"10.1016/j.genrep.2024.102067","url":null,"abstract":"<div><h3>Background</h3><div>Idiopathic pulmonary fibrosis (IPF) is characterized by progressive fibrosis in the lung parenchyma. Given the fact that IPF patients are at significant risk of developing lung cancer (LC), the overlapping gene signatures between IPF and LC need to be explored.</div></div><div><h3>Methods</h3><div>Two datasets (GSE79544 and GSE103888) were procured from the Gene Expression Omnibus repository and used to determine the overlapping genes between IPF and LC. Next, the prediction ability of these genes in differentiating the diseased group from controls was explored using two machine learning (ML) models (random forest and k-nearest neighbor). Potential drugs targeting the candidate genes were identified, and advanced structural analysis was conducted to determine the binding affinity between the candidate drug and target receptor.</div></div><div><h3>Result</h3><div>A total of ten common genes (<em>CCL13</em>, <em>CXCL2</em>, <em>MALT1</em>, <em>MARCKS</em>, <em>PLA2G7</em>, <em>SEMA6B</em>, <em>SFTPB</em>, <em>SPARC</em>, <em>SPP1</em>, and <em>TLCD2</em>) are differentially expressed in IPF and LC as compared to the controls. <em>PLA2G7</em> demonstrated promising potential in differentiating between IPF, LC, and controls. The increased expression correlated with poor survival in patients with LC. The expression of <em>PLA2G7</em> indicated a similar trend in the validation dataset. Darapladib, a selective inhibitor that belongs to toxicity class 4 and lethal dose50 value of 800 mg/kg exhibited maximum potential in targeting <em>PLA2G7</em> with a binding affinity score of −9.2 kcal/mol (chain A) and −9.3 kcal/mol (chain B), respectively.</div></div><div><h3>Conclusion</h3><div>The present study is the first of its kind that combines in-silico and ML algorithms to identify the gene signatures and promising drugs for treating the progression of LC in patients with IPF.</div></div>","PeriodicalId":12673,"journal":{"name":"Gene Reports","volume":null,"pages":null},"PeriodicalIF":1.0,"publicationDate":"2024-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142573010","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}
Gene ReportsPub Date : 2024-10-29DOI: 10.1016/j.genrep.2024.102071
Astrid S. Espinoza García , Rosa L. Díaz Chávez , Elia H. Valdés Miramontes , Isela Parra Rojas , Zyanya Reyes Castillo
{"title":"Genetic variants in ghrelin (rs27647, rs696217) and leptin (rs7799039) are not associated with body composition parameters but are related to appetitive traits in Mexican young adults","authors":"Astrid S. Espinoza García , Rosa L. Díaz Chávez , Elia H. Valdés Miramontes , Isela Parra Rojas , Zyanya Reyes Castillo","doi":"10.1016/j.genrep.2024.102071","DOIUrl":"10.1016/j.genrep.2024.102071","url":null,"abstract":"<div><h3>Purpose</h3><div>Obesity develops as a result of the interplay between obesogenic environment and the individual's eating behaviors. Genetic variants in genes coding for neuropeptides involved in metabolic regulation, such as ghrelin (<em>GHRL</em>) and leptin (<em>LEP</em>) have been associated to development of obesity and its role on appetite regulation has also been suggested. The purpose of this exploratory study was to evaluate the association of genetic variants at <em>GHRL</em> (rs27647, rs696217) and <em>LEP</em> (rs7799039) with body composition parameters as well as food approach and food avoidance appetitive traits in young adults from western Mexico.</div></div><div><h3>Methods</h3><div>255 young adults were included. Body composition parameters were measured by bioelectrical bioimpedance, and appetitive traits were assessed via Adult Eating Behavior Questionnaire (AEBQ). Genotyping of rs27647, rs696217 and rs7799039 variants was performed using PCR-RFLP techniques.</div></div><div><h3>Results and conclusion</h3><div>Median age for both sexes were 20 (19–21) years, mean body fat percentage was 36.1 ± 7.8 for women, and for men 23.7 ± 7.4. The evaluated genetic variants were not associated to body composition parameters, however GG genotype of the rs696217 variant in <em>GHRL</em> was associated to higher scores on food approach appetitive traits (enjoyment of food <em>p</em> < 0.01 and food responsiveness <em>p</em> < 0.0009). In addition, the GG genotype of rs7799039 variant in <em>LEP</em> was associated to higher scores in food avoidance appetitive traits (emotional undereating <em>p</em> < 0.004 and food fussiness <em>p</em> < 0.001). Our results suggest an association between genetic variants in <em>GHRL</em> and <em>LEP</em> genes with appetitive traits in Mexican young adults, showing an indirect genetic link with obesity, through eating behaviors modulation.</div></div>","PeriodicalId":12673,"journal":{"name":"Gene Reports","volume":null,"pages":null},"PeriodicalIF":1.0,"publicationDate":"2024-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142573011","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}
Gene ReportsPub Date : 2024-10-28DOI: 10.1016/j.genrep.2024.102068
Razieh Farkhondeh , Seyedeh Zahra Hasanpour , Mohsen Hamidpour , Mehdi Allah Bakhshian , Seyedeh Ommolbanin Ghasemian , Majid Gholami-Ahangaran
{"title":"The expression of CCL-3, FGF2, TPO and CTGF in newly diagnosed acute myeloblastic leukemia (AML)","authors":"Razieh Farkhondeh , Seyedeh Zahra Hasanpour , Mohsen Hamidpour , Mehdi Allah Bakhshian , Seyedeh Ommolbanin Ghasemian , Majid Gholami-Ahangaran","doi":"10.1016/j.genrep.2024.102068","DOIUrl":"10.1016/j.genrep.2024.102068","url":null,"abstract":"<div><h3>Introduction</h3><div>The interaction between Bone marrow (BM) microenvironment and acute myeloblastic leukemia (AML) cells is an essential event in the development of leukemia.</div></div><div><h3>Material and methods</h3><div>In this study investigated the expression of fibroblastic growth factor 2 (FGF2), chemokine (C<img>C motif) ligand 3 (CCL-3), thrombopoietin (TPO) and connective tissue growth factor (CTGF) in 60 newly diagnosed AML patients and 60 healthy volunteers. BM and peripheral blood samples were collected from patients and healthy individuals and the expression of genes was assessed using qRT-PCR. The obtained data were analyzed using <em>t</em>-test, one-way ANOVA and Pearson correlation coefficient.</div></div><div><h3>Results</h3><div>The results showed that while the expression of CCL-3 and FGF2 was upregulated, the expression of TPO was downregulated in AML patients as compared with the control group. We also failed to find any difference in the expression of CTGF between patients and healthy individuals. Moreover, we found that there was no association between gene expression and the age and gender of AML patients. Only the expression of CTGF had a negative correlation with the percentage of blasts in AML patients. A positive significant correlation between FGF2 and CCL-3, FGF2 and TPO, FGF2 and CTGF as well as CCL-3 and TPO were detected.</div></div><div><h3>Conclusions</h3><div>This study proposed that the expression of growth factors and cytokines could be used as a prognostic factor. However, to gain better insight into the precise role of these factors, further studies at larger statistical population are required.</div></div>","PeriodicalId":12673,"journal":{"name":"Gene Reports","volume":null,"pages":null},"PeriodicalIF":1.0,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142560761","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}
Gene ReportsPub Date : 2024-10-28DOI: 10.1016/j.genrep.2024.102070
Chayna Singha Mahapatra , Aruna Kuniyal , Gaurav Sharma , Shyma K. Latheef , Amit Kumar , Pronab Dhar
{"title":"CRISPR/Cas mediated disruption of the Porcine Circovirus-1 gene present in PK-15 cells using sgRNA cocktail","authors":"Chayna Singha Mahapatra , Aruna Kuniyal , Gaurav Sharma , Shyma K. Latheef , Amit Kumar , Pronab Dhar","doi":"10.1016/j.genrep.2024.102070","DOIUrl":"10.1016/j.genrep.2024.102070","url":null,"abstract":"<div><div>Porcine circovirus 1(PCV-1) is an inherent contaminant in all cells of porcine origin, including cell lines. Porcine Kidney 15 (PK-15) cells are one such cell line which are widely being used for the propagation of different porcine viruses and development of cell culture vaccines for pigs. PCV1 is a single-stranded DNA virus that remain in circular form with a high copy number inside the host cell. The presence of PCV1virus in PK-15 cells may affect the yield of porcine viruses and other vaccine strains like Classical Swine Fever Virus (CSFV) propagated in these cells. Hence the present study was conducted to explore the promising CRISPR/Cas9 tool in cleaving PCV-1 DNA from PK-15 cells, followed by the evaluation of these cells for producing CSF vaccine virus with better yield. PK-15 cells were subjected to CRISPR/Cas9 mediated cleavage with a cocktail of guide RNAs (sgRNAs) to cleave PCV-1 DNA. A significant reduction of PCV-1 DNA in the transfected cells was observed in droplet digital PCR and real-time PCR; indicating successful targeting of PCV-1 DNA by the chosen sgRNAs. Further, the PCV-1 cleaved PK-15 cells were infected with CSFV and found to have a better yield of harvested virus. Based on the present study, it can be suggested that knocking out of PCV-1 DNA from PK-15 cells offers a promising platform for generating contaminant free cell lines and vaccine development with higher yield.</div></div>","PeriodicalId":12673,"journal":{"name":"Gene Reports","volume":null,"pages":null},"PeriodicalIF":1.0,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142573479","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}