Genome engineering using the CRISPR-Cas9 system
نویسندگان
چکیده
منابع مشابه
Optimization of Genome Engineering Approaches with the CRISPR/Cas9 System
Designer nucleases such as TALENS and Cas9 have opened new opportunities to scarlessly edit the mammalian genome. Here we explored several parameters that influence Cas9-mediated scarless genome editing efficiency in murine embryonic stem cells. Optimization of transfection conditions and enriching for transfected cells are critical for efficiently recovering modified clones. Paired gRNAs and w...
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Targeted genome engineering has been instrumental for the study of biological processes, and it holds great promise for the treatment of disease. Historically, gene targeting by homologous recombination has been the preferred method to modify specific genes in mouse and human cells (Fig. 1). However, this approach is hampered by low efficiency, the requirement for drug selection to detect targe...
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Targeted mutagenesis based on homologous recombination has been a powerful tool for understanding the mechanisms underlying development, normal physiology, and disease. A recent breakthrough in genome engineering technology based on the class of RNA-guided endonucleases, such as clustered regularly interspaced short palindromic repeats (CRISPR)-associated Cas9, is further revolutionizing biolog...
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Toxoplasma gondii is a parasite of humans and animals, and a model for other apicomplexans including Plasmodium spp., the causative agents of malaria. Despite many advances, manipulating the T. gondii genome remains labor intensive, and is often restricted to lab-adapted strains or lines carrying mutations that enable selection. Here, we use the RNA-guided Cas9 nuclease to efficiently generate ...
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The ability to modify the murine genome is perhaps one of the most important developments in modern biology. However, traditional methods of genomic engineering are costly and relatively clumsy in their approach. The use of programmable nucleases such as zinc finger nucleases and transcription activator-like effector nucleases significantly improved the precision of genome-editing technology, b...
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ژورنال
عنوان ژورنال: Nature Protocols
سال: 2013
ISSN: 1754-2189,1750-2799
DOI: 10.1038/nprot.2013.143