2022
DOI: 10.17504/protocols.io.81wgb6r5qlpk/v1
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How to select the best gRNA(s) for frameshift knockouts in zebrafish v1

Abstract: Remember to cite the protocol if it is helpful to you! This protocol describes my approach (as of April 2022) to select gRNAs to generate frameshift knockouts in zebrafish using CRISPR-Cas9. It assumes you are selecting 3 gRNAs to generate F0 knockouts, following the method we described in doi: 10.7554/eLife.59683 and protocol 10.17504/protocols.io.bs2rngd6. Are you making a stable knockout line carrying a frameshift mutation? You can also follow this protocol to select a unique gRNA. Get in touch for quest… Show more

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“…We performed F0 knockouts similar as described in Kroll et al 44,67,68 44 ) with pseudo-random sequences predicted to not match any genomic locus. For all experiments scramble RNP injected siblings were used as controls.…”
Section: F0 Knockoutsmentioning
confidence: 99%
“…We performed F0 knockouts similar as described in Kroll et al 44,67,68 44 ) with pseudo-random sequences predicted to not match any genomic locus. For all experiments scramble RNP injected siblings were used as controls.…”
Section: F0 Knockoutsmentioning
confidence: 99%