Knockout Rats Produced Using Designed Zinc Finger Nucleases

By A. Geurts, G. Cost, Yevgeniy Freyvert, B. Zeitler, Jeffrey C. Miller, Vivian M. Choi, Shirin S Jenkins, Adam Wood, X. Cui, Xiangdong Meng and 16 more. Science, 2009

942 citations recorded by the source corpus.

Targeted gene disruption in rats paves the way for new human disease models. The toolbox of rat genetics currently lacks the ability to introduce site-directed, heritable mutations into the genome to create knockout animals. By using engineered zinc-finger nucleases (ZFNs) designed to target an integrated reporter and two endogenous rat genes, Immunoglobulin M (IgM) and Rab38, we demonstrate that a single injection of DNA or messenger RNA encoding ZFNs into the one-cell rat embryo leads to a high frequency of animals carrying 25 to 100% disruption at the target locus. These mutations are faithfully and efficiently transmitted through the germline. Our data demonstrate the feasibility of targeted gene disruption in multiple rat strains within 4 months time, paving the way to a humanized monoclonal antibody platform and additional human disease models.

Subfields: Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/442a39fd945b00a15ff3072714366b40eefb6ca7

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Genetics

In Molecular Biology

Citations in the corpus

Genome Engineering With Zinc-Finger Nucleases (2011, background): “Now embryo injection of ZFN mRNAs offers the prospect of creating targeted mutations in the rat (Geurts et al. 2009; Mashimo et al. 2010).”
TALENs: a widely applicable technology for targeted genome editing (2012, neutral): “…trout 38 , catfish 39 , sea urchin 40 , frog 41 , pig 42 , cattle 43 , cricket 8 , rabbit 44 , silkworm 45 , butterfly 46 , mouse 47, 48 , rat 49, 50 , soybean 51, 52 , thale cress 53, 54 , corn 55 , tobacco 56 , petunia 57 , hamster cells 58 , and human somatic 59, 60 and pluripotent stem cells…”
One-step generation of mice carrying reporter and conditional alleles by CRISPR/Cas mediated genome engineering (2013, positive): “The production of genetically modified mice and rats has been greatly accelerated by novel approaches using direct injection of DNA or mRNA of site-specific nucleases into the one-cell-stage embryo, generating DNA double-strand breaks (DSB) at specified sequences leading to targeted mutations (Carbe”
One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering (2013, neutral): “DSBs induced by these site-specific nucleases can then be repaired by error-prone nonhomologous end joining (NHEJ) resulting in mutant mice and rats carrying deletions or insertions at the cut site (Carbery et al., 2010; Geurts et al., 2009; Sung et al., 2013; Tesson et al., 2011).”
Loss-of-function approaches in comparative physiology: is there a future for knockdown experiments in the era of genome editing? (2019, methodology): “Genome editing using targetable nucleases such as ZFNs (Bibikova et al., 2002; Doyon et al., 2008; Geurts et al., 2009; Urnov et al., 2010), TALENs (Huang et al., 2011; Tesson et al., 2011; Joung and Sander, 2013) and CRISPR/Cas9 (Cong et al., 2013; Jinek et al., 2013; Mali et al., 2013a; Hwang et a”
A novel TALE nuclease scaffold enables high genome editing activity in combination with low toxicity (2011, background): “Accordingly, designer nuclease-based genome engineering has been successfully established in more than 10 model organisms thus far, including plants (3,4), invertebrates (5), fish (6,7) and mammals (8).”
CRISPR/Cas9 systems targeting β-globin and CCR5 genes have substantial off-target activity (2013, background): “For example, engineered zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) have generated hundreds of animal models for disease studies (1,2), and nuclease-based treatment strategies are currently undergoing clinical trials.”
Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity (2013, background): “…DNA nuclease technologies have enabled targeted modification of endogenous genomic sequences with high efficiency, particularly in species that have proven traditionally genetically intractable (Carlson et al., 2012; Geurts et al., 2009; Takasu et al., 2010; Watanabe et al., 2012).”