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

• Published in: Developmental cell Vol. 51; no. 5; pp. 645 - 657.e4
• Main Authors: Hoshijima, Kazuyuki, Jurynec, Michael J., Klatt Shaw, Dana, Jacobi, Ashley M., Behlke, Mark A., Grunwald, David Jonah
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 02.12.2019
• Subjects: CRISPR-Cas9 mutagenesis; deletion mutations; F0 screen; genome editing; targeted mutagenesis; zebrafish genetics; genome editing; F0 screen; deletion mutations; CRISPR-Cas9 mutagenesis; zebrafish genetics; targeted mutagenesis

Inconsistent activity limits the use of CRISPR-Cas9 in zebrafish. We show supernumerary guanine nucleotides at the 5′ ends of single guide RNAs (sgRNAs) account for diminished CRISPR-Cas9 activity in zebrafish embryos. Genomic sequences can be targeted consistently with extremely high efficiency usi...