mRNA processing in mutant zebrafish lines generated by chemical and CRISPR-mediated mutagenesis produces unexpected transcripts that escape nonsense-mediated decay
| العنوان: | mRNA processing in mutant zebrafish lines generated by chemical and CRISPR-mediated mutagenesis produces unexpected transcripts that escape nonsense-mediated decay |
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| المؤلفون: | Catherine M Scahill, Jennifer L. Anderson, Hui Wang, Meng-Chieh Shen, Shao J. Du, Frederick J. Tan, Steven A. Farber, Timothy Mulligan, Elisabeth M. Busch-Nentwich |
| المساهمون: | Anderson, Jennifer L [0000-0002-7015-5791], Mulligan, Timothy S [0000-0002-6046-4523], Wang, Hui [0000-0002-9432-6233], Busch-Nentwich, Elisabeth M [0000-0001-6450-744X], Farber, Steven A [0000-0002-8037-7312], Apollo - University of Cambridge Repository |
| المصدر: | PLoS Genetics PLoS Genetics, Vol 13, Iss 11, p e1007105 (2017) |
| سنة النشر: | 2017 |
| مصطلحات موضوعية: | 0301 basic medicine, Cancer Research, Life Cycles, Embryology, RNA Stability, Nonsense-mediated decay, Mutant, Oligonucleotides, Gene Expression, Artificial Gene Amplification and Extension, Morpholino, medicine.disease_cause, Polymerase Chain Reaction, Biochemistry, Exon, Database and Informatics Methods, Larvae, RNA interference, Genome editing, CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats, Zebrafish, Antisense Oligonucleotides, Genetics (clinical), Genetics, Gene Editing, Mutation, Genome, biology, Nucleotides, Messenger RNA, Fishes, Eukaryota, Nonsense Mutation, Exons, Genomics, Animal Models, Nucleic acids, Phenotypes, Phenotype, Experimental Organism Systems, Genetic interference, Codon, Nonsense, Osteichthyes, Perspective, Vertebrates, Epigenetics, Genetic Engineering, Sequence Analysis, Research Article, Biotechnology, lcsh:QH426-470, Bioinformatics, Mutagenesis (molecular biology technique), Genome Complexity, Research and Analysis Methods, 03 medical and health sciences, Model Organisms, Sequence Motif Analysis, medicine, Point Mutation, Animals, RNA, Messenger, Molecular Biology Techniques, Molecular Biology, Ecology, Evolution, Behavior and Systematics, Embryos, Organisms, Computational Biology, Biology and Life Sciences, biology.organism_classification, Introns, Reverse Genetics, Nonsense Mediated mRNA Decay, lcsh:Genetics, 030104 developmental biology, Fish, Mutagenesis, RNA, Developmental Biology |
| الوصف: | As model organism-based research shifts from forward to reverse genetics approaches, largely due to the ease of genome editing technology, a low frequency of abnormal phenotypes is being observed in lines with mutations predicted to lead to deleterious effects on the encoded protein. In zebrafish, this low frequency is in part explained by compensation by genes of redundant or similar function, often resulting from the additional round of teleost-specific whole genome duplication within vertebrates. Here we offer additional explanations for the low frequency of mutant phenotypes. We analyzed mRNA processing in seven zebrafish lines with mutations expected to disrupt gene function, generated by CRISPR/Cas9 or ENU mutagenesis methods. Five of the seven lines showed evidence of altered mRNA processing: one through a skipped exon that did not lead to a frame shift, one through nonsense-associated splicing that did not lead to a frame shift, and three through the use of cryptic splice sites. These results highlight the need for a methodical analysis of the mRNA produced in mutant lines before making conclusions or embarking on studies that assume loss of function as a result of a given genomic change. Furthermore, recognition of the types of adaptations that can occur may inform the strategies of mutant generation. Author summary The recent rise of reverse genetic, gene targeting methods has allowed researchers to readily generate mutations in any gene of interest with relative ease. Should these mutations have the predicted effect on the mRNA and encoded protein, we would expect many more abnormal phenotypes than are typically being seen in reverse genetic screens. Here we set out to explore some of the reasons for this discrepancy by studying seven separate mutations in zebrafish. We present evidence that thorough cDNA sequence analysis is a key step in assessing the likelihood that a given mutation will produce hypomorphic or null alleles. This study reveals that mRNA processing in the mutant background often produces transcripts that escape nonsense-mediated decay, thereby potentially preserving gene function. By understanding the ways that cells avoid the deleterious consequences of mutations, researchers can better design reverse genetic strategies to increase the likelihood of gene disruption. |
| وصف الملف: | application/pdf |
| تدمد: | 1553-7404 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::763c46cb4449f034c0f08c67681fe848 https://pubmed.ncbi.nlm.nih.gov/29470494 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....763c46cb4449f034c0f08c67681fe848 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15537404 |
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