Publication: Maximizing mutagenesis with solubilized CRISPR-Cas9 ribonucleoprotein complexes
Maximizing mutagenesis with solubilized CRISPR-Cas9 ribonucleoprotein complexes
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Burger, A., Lindsay, H., Felker, A., Hess, C., Anders, C., Chiavacci, E., Zaugg, J., Weber, L. M., Catena, R., Jinek, M., Robinson, M. D., & Mosimann, C. (2016). Maximizing mutagenesis with solubilized CRISPR-Cas9 ribonucleoprotein complexes. Development, 143(11), 2025–2037. https://doi.org/10.1242/dev.134809
Abstract
Abstract
Abstract
CRISPR-Cas9 enables efficient sequence-specific mutagenesis for creating somatic or germline mutants of model organisms. Key constraints in vivo remain the expression and delivery of active Cas9-sgRNA ribonucleoprotein complexes (RNPs) with minimal toxicity, variable mutagenesis efficiencies depending on targeting sequence, and high mutation mosaicism. Here, we apply in vitro assembled, fluorescent Cas9-sgRNA RNPs in solubilizing salt solution to achieve maximal mutagenesis efficiency in zebrafish embryos. MiSeq-based sequence analysi
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Creators (Authors)
- Burger, Alexa
- Lindsay, Helen
- Felker, Anastasia
- Hess, Christopher
- Anders, Carolin
- Chiavacci, Elena
- Zaugg, Jonas
- Weber, Lukas M
- Catena, Raul
- Jinek, Martin
- Mosimann, Christian
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Burger, A., Lindsay, H., Felker, A., Hess, C., Anders, C., Chiavacci, E., Zaugg, J., Weber, L. M., Catena, R., Jinek, M., Robinson, M. D., & Mosimann, C. (2016). Maximizing mutagenesis with solubilized CRISPR-Cas9 ribonucleoprotein complexes. Development, 143(11), 2025–2037. https://doi.org/10.1242/dev.134809
