Chapter title |
Generation of Zebrafish Zebrafish Models by CRISPRCRISPR /Cas9Cas9 Genome Editing Genome Editing.
|
---|---|
Chapter number | 24 |
Book title |
Neuronal Cell Death
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2152-2_24 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2151-5, 978-1-4939-2152-2
|
Authors |
Alexander Hruscha, Bettina Schmid, Hruscha, Alexander, Schmid, Bettina |
Abstract |
The CRISPR CRISPR /CasCas system identified in archaeaarchaea has been adopted and optimized for genomegenome editing purposes in zebrafishzebrafish . In vitroIn vitro transcribed guide RNAguide RNA and Cas9 Cas9 mRNAmRNA are microinjectedmicroinjected into fertilizedfertilized zebrafish embryosembryos to edit the zebrafish genome. Here, we describe how to design a gRNAgRNA , a fast method for in vitroin vitro transcriptiontranscription of gRNA from oligonucleotidesoligonucleotides , microinjectionmicroinjection into fertilized zebrafish embryos, and a PCRPCR -based restriction fragmentrestriction fragment length assayrestriction fragment length assay to identify mutationsmutations at the gRNA target sitetarget site . |
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