Detection of genome-edited mutant clones by a simple competition-based PCR method
نویسندگان
چکیده
منابع مشابه
Detection of genome-edited mutant clones by a simple competition-based PCR method
Genome editing by the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats / CRISPR-associated protein 9) system is a revolutionary strategy to study gene functions. Since the efficiency of gene disruption in cell culture does not reach 100% typically, cloning of mutant cells is often performed to obtain fully mutated cells. Therefore, a method to discriminate accurately mutat...
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Screening or selection of mutants is an essential step in genetics. Mutants are invaluable tools for the elucidation of biochemical pathways, for defining the roles of genetic units in a particular pathway and for understanding the relationship between enzyme structure and function (3). For mutant searches, there are two approaches: (i) positive selection for mutants that can grow under selecti...
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The rapid adoption of gene editing tools such as CRISPRs and TALENs for research and eventually therapeutics necessitates assays that can rapidly detect and quantitate the desired alterations. Currently, the most commonly used assay employs "mismatch nucleases" T7E1 or "Surveyor" that recognize and cleave heteroduplexed DNA amplicons containing mismatched base-pairs. However, this assay is pron...
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Successful production of genome-edited rats by the rGONAD method
BACKGROUND Recent progress in development of the CRISPR/Cas9 system has been shown to be an efficient gene-editing technology in various organisms. We recently developed a novel method called Genome-editing via Oviductal Nucleic Acids Delivery (GONAD) in mice; a novel in vivo genome editing system that does not require ex vivo handling of embryos, and this technology is newly developed and rena...
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ژورنال
عنوان ژورنال: PLOS ONE
سال: 2017
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0179165