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Cited by 260

The DOI matches its Crossref record; 10 of the 10 references it cites were found in a registry or an index.

Highly Efficient CRISPR-Cas9-Based Methods for Generating Deletion Mutations and F0 Embryos that Lack Gene Function in Zebrafish

Open access
Hoshijima, Kazuyuki, Jurynec, Michael J., Klatt Shaw, Dana, Jacobi, Ashley M., Behlke, Mark A., Grunwald, David Jonah·2019·Developmental Cell
DOI 10.1016/j.devcel.2019.10.004
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Abstract

SUMMARY Inconsistent activity limits use of CRISPR/Cas9 in zebrafish. We show supernumerary guanine nucleotides at the 5’ ends of sgRNAs account for diminished CRISPR/Cas9 activity in zebrafish embryos. Genomic sequences can be targeted consistently with extremely high efficiency using Cas9 RNPs containing either a sgRNA molecule or a synthetic crRNA:tracrRNA duplex that perfectly matches the protospacer target site. Following injection of zebrafish eggs with such RNPs, virtually every copy of a targeted locus harbors an induced indel mutation. Loss of gene function is often complete, as F0 embryos closely resemble true null mutants without detectable non-specific effects. Mosaicism is sufficiently low in F0 embryos that cell non-autonomous gene functions can be probed effectively and redundant activities of genes can be uncovered when two genes are targeted simultaneously. Finally, heritable deletion mutations of at least 50 kbp can be readily induced using pairs of duplex guide RNPs targeted to a single chromosome.

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Metadata

Authors
Hoshijima, K., Jurynec, M. J., Klatt Shaw, D., Jacobi, A. M., Behlke, M. A., & Grunwald, D. J.
Venue
Developmental Cell
Publisher
Elsevier BV
Published
2019-12
Volume
51
Issue
5
Pages
645-657.e4
ISSN
1534-5807 (ISSN-L)
DOI
10.1016/j.devcel.2019.10.004
OpenAlex
W2987688991
Cited by
260
References
10
FWCI
7.81
Type
journal article
Language
en
Open access
bronze

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