Scarless genome editing: progress towards understanding genotype–phenotype relationships
نویسندگان
چکیده
منابع مشابه
Optimization of scarless human stem cell genome editing
Efficient strategies for precise genome editing in human-induced pluripotent cells (hiPSCs) will enable sophisticated genome engineering for research and clinical purposes. The development of programmable sequence-specific nucleases such as Transcription Activator-Like Effectors Nucleases (TALENs) and Cas9-gRNA allows genetic modifications to be made more efficiently at targeted sites of intere...
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Ali, M. A. 2014. Understanding Cancer Mutations by Genome Editing. Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine 1054. 37 pp. Uppsala: Acta Universitatis Upsaliensis. ISBN 978-91-554-9106-2. Mutational analyses of cancer genomes have identified novel candidate cancer genes with hitherto unknown function in cancer. To enable phenotyping of mutations in suc...
متن کاملScarless Genome Editing of Human Pluripotent Stem Cells via Transient Puromycin Selection
Genome-edited human pluripotent stem cells (hPSCs) have broad applications in disease modeling, drug discovery, and regenerative medicine. We present and characterize a robust method for rapid, scarless introduction or correction of disease-associated variants in hPSCs using CRISPR/Cas9. Utilizing non-integrated plasmid vectors that express a puromycin N-acetyl-transferase (PAC) gene, whose exp...
متن کاملA versatile and highly efficient method for scarless genome editing in Escherichia coli and Salmonella enterica
BACKGROUND Recently developed methods for genome editing in bacteria take advantage of the introduction of double-strand breaks by I-SceI in a mutation cassette to select for cells in which homologous recombination has healed the break and introduced a desired mutation. This elegantly designed method did not work well in our hands for most genes. RESULTS We corrected a mutation in the gene en...
متن کاملThe no-SCAR (Scarless Cas9 Assisted Recombineering) system for genome editing in Escherichia coli
Genome engineering methods in E. coli allow for easy to perform manipulations of the chromosome in vivo with the assistance of the λ-Red recombinase system. These methods generally rely on the insertion of an antibiotic resistance cassette followed by removal of the same cassette, resulting in a two-step procedure for genomic manipulations. Here we describe a method and plasmid system that can ...
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ژورنال
عنوان ژورنال: Current Genetics
سال: 2018
ISSN: 0172-8083,1432-0983
DOI: 10.1007/s00294-018-0850-8