The DOI matches its Crossref record; 4 of the 4 references it cites were found in a registry or an index.
Scalable and Versatile Genome Editing Using Linear DNAs with Microhomology to Cas9 Sites in Caenorhabditis elegans
Open accessAbstract
Homology-directed repair (HDR) of double-strand DNA breaks is a promising method for genome editing, but is thought to be less efficient than error-prone nonhomologous end joining in most cell types. We have investigated HDR of double-strand breaks induced by CRISPR-associated protein 9 (Cas9) in Caenorhabditis elegans. We find that HDR is very robust in the C. elegans germline. Linear repair templates with short (∼30–60 bases) homology arms support the integration of base and gene-sized edits with high efficiency, bypassing the need for selection. Based on these findings, we developed a systematic method to mutate, tag, or delete any gene in the C. elegans genome without the use of co-integrated markers or long homology arms. We generated 23 unique edits at 11 genes, including premature stops, whole-gene deletions, and protein fusions to antigenic peptides and GFP. Whole-genome sequencing of five edited strains revealed the presence of passenger variants, but no mutations at predicted off-target sites. The method is scalable for multi-gene editing projects and could be applied to other animals with an accessible germline.
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Cites
A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J. A., & Charpentier, E. (2012) · Science · doi:10.1126/science.1225829 · Cited by 17,116
CRISPR-Cas systems for editing, regulating and targeting genomes
Sander, J. D., & Joung, J. K. (2014) · Nature Biotechnology · doi:10.1038/nbt.2842 · Cited by 3,153 · cited 2×
“In many systems, HDR is thought to be less efficient than NHEJ, requiring high concentrations of donor molecules or long homology arms to stimulate recombination (see Beumer et al. 2013 and Sander and Joung 2014 for review).”
BackgroundYang, H., Wang, H., Shivalila, C. S., Cheng, A. W., Shi, L., & Jaenisch, R. (2013) · Cell · doi:10.1016/j.cell.2013.08.022 · Cited by 1,596
Optimized CRISPR/Cas tools for efficient germline and somatic genome engineering in Drosophila
Port, F., Chen, H.-M., Lee, T., & Bullock, S. L. (2014) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1405500111 · Cited by 1,046
Cited by
Rapid protein depletion in human cells by auxin-inducible degron tagging with short homology donors
Natsume, T., Kiyomitsu, T., Saga, Y., & Kanemaki, M. T. (2016) · Cell Reports · doi:10.1016/j.celrep.2016.03.001 · Cited by 673
Paix, A., Folkmann, A., Rasoloson, D., & Seydoux, G. (2015) · Genetics · doi:10.1534/genetics.115.179382 · Cited by 675
Self-citationZhang, L., Ward, J. D., Cheng, Z., & Dernburg, A. F. (2015) · Development · doi:10.1242/dev.129635 · Cited by 599
Streamlined Genome Engineering with a Self-Excising Drug Selection Cassette
Dickinson, D. J., Pani, A. M., Heppert, J. K., Higgins, C. D., & Goldstein, B. (2015) · Genetics · doi:10.1534/genetics.115.178335 · Cited by 586
Wang, J. T., Smith, J., Chen, B.-C., Schmidt, H., Rasoloson, D., Paix, A., Lambrus, B. G., Calidas, D., Betzig, E., & Seydoux, G. (2014) · eLife · doi:10.7554/elife.04591 · Cited by 364
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Metadata
- Authors
- Paix, A., Wang, Y., Smith, H. E., Lee, C.-Y. S., Calidas, D., Lu, T., Smith, J., Schmidt, H., Krause, M. W., & Seydoux, G.
- Venue
- Genetics
- Publisher
- Oxford University Press (OUP)
- Published
- 2014-09-23
- Volume
- 198
- Issue
- 4
- Pages
- 1347-1356
- ISSN
- 0016-6731 (ISSN-L)
- 1943-2631
- OpenAlex
- W2171730432
- Cited by
- 282
- References
- 4
- FWCI
- 9.73
- Type
- journal article
- Language
- en
- Open access
- green
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- Paix, A., Wang, Y., Smith, H. E., Lee, C.-Y. S., Calidas, D., Lu, T., Smith, J., Schmidt, H., Krause, M. W., & Seydoux, G. (2014). Scalable and Versatile Genome Editing Using Linear DNAs with Microhomology to Cas9 Sites in Caenorhabditis elegans. Genetics. https://doi.org/10.1534/genetics.114.170423
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