The DOI matches its Crossref record; 4 of the 4 references it cites were found in a registry or an index.
Validation of microinjection methods for generating knockout mice by CRISPR/Cas-mediated genome engineering
Abstract
The CRISPR/Cas system, in which the Cas9 endonuclease and a guide RNA complementary to the target are sufficient for RNA-guided cleavage of the target DNA, is a powerful new approach recently developed for targeted gene disruption in various animal models. However, there is little verification of microinjection methods for generating knockout mice using this approach. Here, we report the verification of microinjection methods of the CRISPR/Cas system. We compared three methods for injection: (1) injection of DNA into the pronucleus, (2) injection of RNA into the pronucleus and (3) injection of RNA into the cytoplasm. We found that injection of RNA into the cytoplasm was the most efficient method in terms of the numbers of viable blastocyst stage embryos and full-term pups generated. This method also showed the best overall knockout efficiency.
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“Traditional physical methods such as microinjection [136,137] or electroporation [138,139] have been successfully used with CRISPR to engineer embryonic stem cells and zygotes that later originate genetically modified animals.”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [137]
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“In addition, there is evidence that injection of Cas9 mRNA and sgRNA into the zygote cytoplasm is the most efficient method for yielding normal embryos and full-term mouse pups harboring the desired modification (Horii et al., 2014).”
From Delivering CRISPR: a review of the challenges and approaches (2018) · cites this paper as Horii et al., 2014
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Cites
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Cited by
Sorted by citations
Delivering CRISPR: a review of the challenges and approaches
Lino, C. A., Harper, J. C., Carney, J. P., & Timlin, J. A. (2018) · Drug Delivery · doi:10.1080/10717544.2018.1474964 · Cited by 1,185
Delivery strategies of the CRISPR-Cas9 gene-editing system for therapeutic applications
Liu, C., Zhang, L., Liu, H., & Cheng, K. (2017) · Journal of Controlled Release · doi:10.1016/j.jconrel.2017.09.012 · Cited by 607
Past, present, and future of CRISPR genome editing technologies
Pacesa, M., Pelea, O., & Jinek, M. (2024) · Cell · doi:10.1016/j.cell.2024.01.042 · Cited by 609
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Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes
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Metadata
- Authors
- Horii, T., Arai, Y., Yamazaki, M., Morita, S., Kimura, M., Itoh, M., Abe, Y., & Hatada, I.
- Venue
- Scientific Reports
- Published
- 2014-03-28
- Volume
- 4
- Issue
- 1
- ISSN
- 2045-2322
- PMC
- PMC5380110
- Cited by
- 175
- References
- 4
- Language
- en
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