CRIMP: A CRISPR/Cas9 Insertional Mutagenesis Protocol and the CRIMP Toolkit
Lee B. MilesiD, Vanessa CalcinottoiD, Carmen SonntagiD, Clara LeeiD, Robert J. Bryson‐RichardsoniD
Abstract
We developed a highly efficient targeted insertional mutagenesis system, CRIMP, and an associated plasmid toolkit, CRIMPkit, that disrupts native gene expression by inducing complete transcriptional termination to produce null mutant alleles without inducing genetic compensation. The CRIMPkit contains over 30 ready-to-use plasmid vectors allowing easy and complete mutagenesis of any gene in any reading frame without requiring custom sequences, modification, or subcloning, and also provides a fluorescent readout of successfully mutagenised fish.
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