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The CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (CRISPR-associated nuclease 9) technology enables rapid, targeted, and efficient changes in the genomes of various model organisms. The short guide RNAs (gRNAs) of the CRISPR/Cas9 system can be designed to recognize target DNA within coding regions for functional gene knockouts. Several studies have demonstrated that the CRISPR/Cas9 system efficiently and specifically targets sea urchin genes and results in expected mutant phenotypes. In addition to disrupting gene functions, modifications and additions to the Cas9 protein enable alternative activities targeted to specific sites within the genome. This includes a fusion of cytidine deaminase to Cas9 (Cas9-DA) for single nucleotide conversion in targeted sites. In this chapter, we describe detailed methods for the CRISPR/Cas9 application in sea urchin embryos, including gRNA design, in vitro synthesis of single guide RNA (sgRNA), and the usages of the CRISPR/Cas9 technology for gene knockout and single nucleotide editing. Methods for genotyping the resultant embryos are also provided for assessing efficiencies of gene editing.
Anderson,
mRNA processing in mutant zebrafish lines generated by chemical and CRISPR-mediated mutagenesis produces unexpected transcripts that escape nonsense-mediated decay.
2017, Pubmed
Anderson,
mRNA processing in mutant zebrafish lines generated by chemical and CRISPR-mediated mutagenesis produces unexpected transcripts that escape nonsense-mediated decay.
2017,
Pubmed
Angerer,
Disruption of gene function using antisense morpholinos.
2004,
Pubmed
Calestani,
Isolation of pigment cell specific genes in the sea urchin embryo by differential macroarray screening.
2003,
Pubmed
,
Echinobase
Cameron,
SpBase: the sea urchin genome database and web site.
2009,
Pubmed
,
Echinobase
Cui,
Recent advances in functional perturbation and genome editing techniques in studying sea urchin development.
2017,
Pubmed
,
Echinobase
El-Brolosy,
Genetic compensation: A phenomenon in search of mechanisms.
2017,
Pubmed
Gilbert,
CRISPR-mediated modular RNA-guided regulation of transcription in eukaryotes.
2013,
Pubmed
Hilton,
Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers.
2015,
Pubmed
Jinek,
A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.
2012,
Pubmed
Komor,
Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage.
2016,
Pubmed
Kudtarkar,
Echinobase: an expanding resource for echinoderm genomic information.
2017,
Pubmed
Labun,
CHOPCHOP v2: a web tool for the next generation of CRISPR genome engineering.
2016,
Pubmed
Larson,
CRISPR interference (CRISPRi) for sequence-specific control of gene expression.
2013,
Pubmed
Lin,
Genome editing in sea urchin embryos by using a CRISPR/Cas9 system.
2016,
Pubmed
,
Echinobase
Maeder,
CRISPR RNA-guided activation of endogenous human genes.
2013,
Pubmed
Mellott,
Notch signaling patterns neurogenic ectoderm and regulates the asymmetric division of neural progenitors in sea urchin embryos.
2017,
Pubmed
,
Echinobase
Moreno-Mateos,
CRISPRscan: designing highly efficient sgRNAs for CRISPR-Cas9 targeting in vivo.
2015,
Pubmed
Nakayama,
Simple and efficient CRISPR/Cas9-mediated targeted mutagenesis in Xenopus tropicalis.
2013,
Pubmed
Nishida,
Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems.
2016,
Pubmed
Oulhen,
Transient translational quiescence in primordial germ cells.
2017,
Pubmed
,
Echinobase
Oulhen,
Albinism as a visual, in vivo guide for CRISPR/Cas9 functionality in the sea urchin embryo.
2016,
Pubmed
,
Echinobase
Ran,
Genome engineering using the CRISPR-Cas9 system.
2013,
Pubmed
Rossi,
Genetic compensation induced by deleterious mutations but not gene knockdowns.
2015,
Pubmed
Shevidi,
Single nucleotide editing without DNA cleavage using CRISPR/Cas9-deaminase in the sea urchin embryo.
2017,
Pubmed
,
Echinobase
Stainier,
Guidelines for morpholino use in zebrafish.
2017,
Pubmed
Vejnar,
Optimized CRISPR-Cas9 System for Genome Editing in Zebrafish.
2016,
Pubmed
