CRISPR

The Challenge of Getting CRISPR-based Knock-ins to Work in Zebrafish

Just the other day, I was talking with a collaborator who told me that they didn’t really believe that knock-ins really worked in zebrafish. Although there have been literature reports for years, the number of labs that struggle and fail with this technique far outweigh the number that find success and publish. So I can …

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6 things that can go wrong while making your Zebrafish CRISPR Knock-In

With the advent of targeted nucleases like zinc-finger nucleases (ZFNs), TALE nucleases (TALENs), and clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas), zebrafish researchers were able to begin interrogating gene function and regions of interest with a precision that was previously out of reach. With these innovations, researchers could not only disrupt …

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Zebrafish modeling for the clinic: Rapid in vivo functional testing of patient variants for clinical applications.

We use the zebrafish (Danio rerio) as an in vivo model to measure the functional effects of patient-derived genetic variation.  In this way, human genetic variants identified in the clinic are quantitatively and qualitatively connected to model animal phenotypes.  As a proof of concept study, we used precision gene editing and transient knockdown approaches targeting stxbp1a – the …

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CRISPR technologies enable humanized animal models to aid disease research

C. elegans as a model to evaluate the function of disease genes In 1998, the soil nematode Caenorhabditis elegans became the first multicellular organism of which the genome has been sequenced completely [1]. One surprising result of this approach was that ~65% of the human disease genes have a counterpart in the worm [2]. In …

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