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Browsing by Author "Cassidy, Annelise"
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Item CRISPR-Cas9-mediated insertion of a short artificial intron for the generation of conditional alleles in mice(Elsevier, 2023) Cassidy, Annelise; Pelletier, Stephane; Medical and Molecular Genetics, School of MedicineIn this protocol, we describe the generation of conditional alleles in mice using the DECAI (DEgradation based on Cre-regulated Artificial Intron) approach. We detail steps for the CRISPR-mediated insertion of the short DECAI cassette within exon 3 of Scyl1 and the functional validation of alleles at genomic, transcriptomic, and protein levels. This strategy simplifies the process of generating mice with conditional alleles.Item Novel methods for the generation of genetically engineered animal models(Elsevier, 2023) Cassidy, Annelise; Onal, Melda; Pelletier, Stephane; Medical and Molecular Genetics, School of MedicineGenetically modified mouse models have shaped our understanding of biological systems in both physiological and pathological conditions. For decades, mouse genome engineering has relied on transgenesis and spontaneous gene replacement in embryonic stem (ES) cells. While these technologies provided a wealth of knowledge, they remain imprecise and expensive to use. Recent advances in genome editing technologies such as the development of targetable nucleases, the improvement of delivery systems, and the simplification of targeting strategies now allow for the rapid, precise manipulation of the mouse genome. In this review article, we discuss novel methods and targeting strategies for the generation of mouse models for the study of bone and skeletal muscle biology.