Heritable genome editing in C. elegans via a CRISPR-Cas9 system

By Ari E. Friedland, Yonatan B. Tzur, Kevin M. Esvelt, M. Colaiácovo, George M. Church, John A. Calarco. Nature Methods, 2013

928 citations recorded by the source corpus.

We report the use of clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated endonuclease Cas9 to target genomic sequences in the Caenorhabditis elegans germ line using single-guide RNAs that are expressed from a U6 small nuclear RNA promoter. Our results demonstrate that targeted, heritable genetic alterations can be achieved in C. elegans, providing a convenient and effective approach for generating loss-of-function mutants.

Subfields: Aging, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/3468ffcf34768b7df39e95662f216a3712cf7ed4

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

In Aging

1. Hans Clevers

In Molecular Biology

1. Feng Zhang
2. Jin-Soo Kim
3. Sojung Kim
4. P. Hsu
5. Hans Clevers
6. Jonathan Gootenberg
7. Jennifer A. Doudna
8. S. Cho

Citations in the corpus

• Efficient Marker-Free Recovery of Custom Genetic Modifications with CRISPR/Cas9 in Caenorhabditis elegans (2014, background): “…have recently revolutionized genome editing in a wide range of organisms (Wang et al. 1999; Chiu et al. 2013; Cho et al. 2013; Dicarlo et al. 2013; Friedland et al. 2013; Gratz et al. 2013; Hwang et al. 2013; Jiang et al. 2013; Katic and Großhans 2013; Li et al. 2013; Lo et al. 2013; Nekrasov et…”
• A CRISPR/Cas9 toolkit for multiplex genome editing in plants (2014, background): “The CRISPR/Cas system has been harnessed to achieve efficient genome editing in a variety of organisms, including bacteria, yeast, plants, and animals, as well as human cell lines [12-27].”
• Efficient Mutagenesis by Cas9 Protein-Mediated Oligonucleotide Insertion and Large-Scale Assessment of Single-Guide RNAs (2014, background): “This simplified system has been implemented in a variety of in vivo settings, resulting in efficient mutagenesis by NHEJ-mediated small insertions or deletions [5–9].”
• Gene disruption by cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA (2014, background): “…that (1) the efficiency (on-target effect) and specificity (off-target effect) are strongly affected by Cas9 protein and guide RNA concentrations (Fu et al. 2013; Hsu et al. 2013; Pattanayak et al. 2013), and (2) it is challenging to exactly control these concentrations so that they are the same…”
• Functional repair of CFTR by CRISPR/Cas9 in intestinal stem cell organoids of cystic fibrosis patients. (2013, methodology): “Recently, several groups have demonstrated the use of the CRISPR/Cas9 system for genome engineering in various species (Chang et al., 2013; Cho et al., 2013; Cong et al., 2013; Friedland et al., 2013; Hou et al., 2013; Hwang et al., 2013; Jinek et al., 2013; Li et al., 2013; Mali et al., 2013; Nekra”
• Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins (2014, methodology): “…then, RGENs have been successfully used to modify genomes in various species including model organisms (Cho et al. 2013b; Dickinson et al. 2013; Friedland et al. 2013; Gratz et al. 2013; Li et al. 2013a,c; Wang et al. 2013; Sung et al. 2014) and plants (Li et al. 2013b; Nekrasov et al. 2013;…”
• CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes (2015, background): “…genome editing, gene function investigation, and gene therapy in animals and human cells (Chang et al., 2013; Cho et al., 2013; Cong et al., 2013; Friedland et al., 2013; Hsu et al., 2014; Hwang et al., 2013; Ikmi et al., 2014; Irion et al., 2014; Jinek et al., 2013; Li et al., 2013a; Li et al.,…”