Highly Efficient CRISPR-Cas9-Based Methods for Generating Deletion Mutations and F0 Embryos that Lack Gene Function in Zebrafish
Kazuyuki Hoshijima,Michael J. Jurynec,Dana Klatt Shaw,Ashley M. Jacobi,Mark A. Behlke,David Grunwald +5 more
TLDR
Supernumerary guanine nucleotides at the 5' ends of single guide RNAs (sgRNAs) account for diminished CRISPR-Cas9 activity in zebrafish embryos and heritable deletion mutations of at least 50 kbp can be readily induced using pairs of duplex guide RNPs targeted to a single chromosome.About:
This article is published in Developmental Cell.The article was published on 2019-12-02 and is currently open access. It has received 166 citations till now. The article focuses on the topics: Trans-activating crRNA & CRISPR.read more
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Genetic compensation induced by deleterious mutations but not gene knockdowns
TL;DR: In this article, the authors show that egfl7 mutants do not show any obvious phenotypes while animals injected with egfl 7 morpholino (morphants) exhibit severe vascular defects, indicating that the activation of a compensatory network to buffer against deleterious mutations was not observed after translational or transcriptional knockdown.
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A simple and effective F0 knockout method for rapid screening of behaviour and other complex phenotypes.
François Kroll,Gareth T. Powell,Marcus Ghosh,Gaia Gestri,Paride Antinucci,Timothy J Hearn,Hande Tunbak,Sumi Lim,Harvey W Dennis,Joseph M Fernandez,David Whitmore,David Whitmore,Elena Dreosti,Stephen W. Wilson,Ellen J. Hoffman,Jason Rihel +15 more
TL;DR: This work developed a simple sequencing-free tool to validate gRNAs and a highly effective CRISPR-Cas9 method capable of converting >90% of injected embryos directly into F0 biallelic knockouts, and demonstrates that F0 knockouts reliably recapitulate complex mutant phenotypes.
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Evolutionary conservation, translational relevance and cognitive function: The future of zebrafish in behavioral neuroscience
TL;DR: This review, based upon an Outstanding Achievement Award lecture presented by the author at an IBNS conference, explores the potential reasons for the popularity of the zebrafish and discusses theoretical questions and empirical examples from the author's own laboratory.
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Optic cup morphogenesis requires neural crest-mediated basement membrane assembly
TL;DR: Using multidimensional imaging and computational analyses in zebrafish, it is established that cell movements in the developing optic cup require neural crest, and that optic cup morphogenesis is dependent upon neural crest-derived nidogens, which modulate extracellular matrix assembly.
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Dissecting cell identity via network inference and in silico gene perturbation
TL;DR: In this paper , a machine-learning-based approach, called CellOracle, is applied to well-established paradigms-mouse and human haematopoiesis, and zebrafish embryogenesis-and correctly model reported changes in phenotype that occur as a result of transcription factor perturbation.
References
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