The DOI matches its Crossref record; 3 of the 3 references it cites were found in a registry or an index.
High Efficiency, Homology-Directed Genome Editing in Caenorhabditis elegans Using CRISPR-Cas9 Ribonucleoprotein Complexes
Open accessAbstract
Homology-directed repair (HDR) of breaks induced by the RNA-programmed nuclease Cas9 has become a popular method for genome editing in several organisms. Most HDR protocols rely on plasmid-based expression of Cas9 and the gene-specific guide RNAs. Here we report that direct injection of in vitro–assembled Cas9-CRISPR RNA (crRNA) trans-activating crRNA (tracrRNA) ribonucleoprotein complexes into the gonad of Caenorhabditis elegans yields HDR edits at a high frequency. Building on our earlier finding that PCR fragments with 35-base homology are efficient repair templates, we developed an entirely cloning-free protocol for the generation of seamless HDR edits without selection. Combined with the co-CRISPR method, this protocol is sufficiently robust for use with low-efficiency guide RNAs and to generate complex edits, including ORF replacement and simultaneous tagging of two genes with fluorescent proteins.
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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
The new frontier of genome engineering with CRISPR-Cas9
Doudna, J. A., & Charpentier, E. (2014) · Science · doi:10.1126/science.1258096 · Cited by 7,062
CRISPR RNA Maturation by Trans-Encoded Small RNA and Host Factor RNase III
Deltcheva, E., Chylinski, K., Sharma, C. M., Gonzales, K., Chao, Y., Pirzada, Z. A., Eckert, M. R., Vogel, J., & Charpentier, E. (2011) · Nature · doi:10.1038/nature09886 · Cited by 2,563
Cited by
Applications of CRISPR technologies in research and beyond
Barrangou, R., & Doudna, J. A. (2016) · Nature Biotechnology · doi:10.1038/nbt.3659 · Cited by 1,005
Wang, H.-X., Li, M., Lee, C. M., Chakraborty, S., Kim, H.-W., Bao, G., & Leong, K. W. (2017) · Chemical Reviews · doi:10.1021/acs.chemrev.6b00799 · Cited by 560
Genome-Editing Technologies: Principles and Applications
Gaj, T., Sirk, S. J., Shui, S.-L., & Liu, J. (2016) · Cold Spring Harbor Perspectives in Biology · doi:10.1101/cshperspect.a023754 · Cited by 381
Kitchen, S., Ratnappan, R., Han, S., Leasure, C., Grill, E., Iqbal, Z., Granger, O., O'Halloran, D. M., & Hawdon, J. M. (2019) · International Journal for Parasitology · doi:10.1016/j.ijpara.2018.12.004 · Cited by 105
Nuclear envelope assembly relies on CHMP-7 in the absence of BAF–LEM-mediated hole closure
Barger, S. R., Penfield, L., & Bahmanyar, S. (2023) · Journal of Cell Science · doi:10.1242/jcs.261385 · Cited by 17
Metadata
- Authors
- Paix, A., Folkmann, A., Rasoloson, D., & Seydoux, G.
- Venue
- Genetics
- Publisher
- Oxford University Press (OUP)
- Published
- 2015-07-17
- Volume
- 201
- Issue
- 1
- Pages
- 47-54
- ISSN
- 0016-6731 (ISSN-L)
- 1943-2631
- OpenAlex
- W2120998017
- Cited by
- 675
- References
- 3
- FWCI
- 17.24
- Type
- journal article
- Language
- en
- Open access
- green
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