Abstract
The clustered, regularly interspaced, short, palindromic repeat (CRISPR)-associated (CAS) nuclease Cas9 has been used in many organisms to generate specific mutations and transgene insertions. Here we describe a method using the S. pyogenes Cas9 in C. elegans that provides a convenient and effective approach for making heritable changes to the worm genome.
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Acknowledgements
We would like to thank the many investigators who have contributed to the recent developments in genome editing and transgenesis techniques in C. elegans. These studies have been referenced throughout our manuscript and should be consulted for additional advice and tips for genome editing. In particular, we thank the Jorgensen, Dupuy, and Goldstein laboratories for the development of highly useful selectable markers and strategies that are effective for both Mos- and CRISPR/Cas9-based editing approaches. Many of the approaches and reagents described by these groups have been helpful in establishing the current methodology described here. We have also greatly appreciated excellent collaborations with Yonatan Tzur, Monica Colaiacovo, Kevin Esvelt, and George Church which led to the original protocol that the current approach is based upon. We also thank Heesun Kim, Masaki Shirayama, and Craig Mello for continued stimulating discussions regarding improvements in this exciting technology. Our work has been supported by US National Institutes of Health Early Independence Award (1DP5OD009153) and additional support from Harvard University to J.A.C. and a Ralph Ellison/American Federation for Aging Research postdoctoral fellowship to A.E.F.
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Calarco, J.A., Friedland, A.E. (2015). Creating Genome Modifications in C. elegans Using the CRISPR/Cas9 System. In: Biron, D., Haspel, G. (eds) C. elegans. Methods in Molecular Biology, vol 1327. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-4939-2842-2_6
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DOI: https://doi.org/10.1007/978-1-4939-2842-2_6
Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-4939-2842-2
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