Monthly Archives: August 2014

Genome Editing: Which Should I Choose, TALEN or CRISPR?

  Ed Davis, Ph.D. Introduction Genome Editing-the ability to make specific changes at targeted genomic sites-is of fundamental importance to researchers in biology and medicine (Bogdanove & Voytas, 2011; van der Oost, et al.,2013). Two genome editing technologies have emerged recently that exploit bacterial systems for plant pathogenesis or adaptive immunity: TALEN (Transcription Activator-Like Effector Nucleases) and CRISPR (Clustered, Regularly Interspaced, Short Palindromic Repeats), respectively. Both TALEN and CRISPR use endonucleases tha......
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Genome Editing in Mammalian Cells: What Do I Do Next?

  Ed Davis, Ph.D. Genome Editing-the ability to make specific changes at targeted genomic sites-is of fundamental importance in biology and medicine (for reviews, see Bogdanove & Voytas, 2011; van der Oost, et al.,2013). Two genome editing technologies have emerged recently that exploit bacterial systems for plant pathogenesis or adaptive immunity: TALEN (Transcription Activator-Like Effector Nucleases) and CRISPR (Clustered, Regularly Interspaced, Short Palindromic Repeats), respectively. Both TALEN and CRISPR use endonucleases that initiate double-strand ......
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CRISPR-Cas9 Specificity: Taming Off-target Mutagenesis

  Ed Davis, Ph.D. Abstract Genome Editing-the ability to make specific changes at targeted genomic sites in complex organisms-is of fundamental importance in biology and medicine (Bogdanove & Voytas, 2011; van der Oost, et al.,2013). Recently, the CRISPR (Clustered, Regularly Interspaced, Short Palindromic Repeats)-Cas (CRISPR- associated) system has become popular for applications such as gene knockouts, making precise, defined base changes, and for transgenesis, to name a few. The ease of design, high efficiency, and relatively low cost of CRISPR-Cas off......
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Genome Editing Technology From GeneCopoeia, Inc.

  Ed Davis, Ph.D. Abstract Genome Editing-the ability to make specific changes at targeted genomic sites-is fundamentally important to researchers in biology and medicine (Bogdanove & Voytas, 2011; van der Oost, et al., 2013). Two genome editing technologies have emerged recently that exploit bacterial systems for plant pathogenesis or adaptive immunity: TALEN (Transcription Activator-Like Effector Nucleases) and CRISPR (Clustered, Regularly Interspaced, Short Palindromic Repeats), respectively. Both TALEN and CRISPR use endonucleases that ini......
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Knockout by TALEN or CRISPR vs. Knockdown by shRNA or siRNA

  Ed Davis, Ph.D. Recent advances in technologies for genome editing-the use of TALEN or CRISPR to make targeted, permanent changes to genes-have revolutionized molecular genetics. They have also presented users with a choice between these relatively new technologies and that of the more established method of RNA interference (RNAi)-mediated knockdown using short hairpin RNA (shRNA) or short interfering RNA (siRNA). In this Technical Note, we explore the differences between the two methods for ablating gene function, and situations where one technology is more ......
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GeneCopoeia Genome Editing Tools for Safe Harbor Integration in Mice and Humans

  Ed Davis, Liuqing Qian, Ruiqing lI, Junsheng Zhou, and Jinkuo Zhang Introduction The ability to introduce transgenes into biological systems, such as cultured cells or animals, is of fundamental importance in biology and medicine. Applications for the introduction of such transgenes include, but are not limited to: 1) Overexpression of a protein of interest in order to detect a phenotype; 2) Tagging of a protein with a fluorescent marker for tracking its localization in cells; 3) Rescue of a mutant phenotype with the wild type allele; and 4) Exp......
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