Design CRISPR-Cas9 Guide RNA and Donor Oligo to Introduce Pathogenic Mutation into Mice
Journal: Journal of Clinical Medicine Research DOI: 10.32629/jcmr.v2i3.459
Abstract
CRISPR-Ca9 system is a newly developed gene-editing technology, which is widely used in biology and medical research. In this project, we want to knock-in a mutation found in a human pedigree into mice through CRISPR-Cas9 technology to validate its pathogenic effect. We download corresponding mice genomic sequences and design guide RNA and donor oligo sequences according to CRISPR-Cas9 target principles. Following experiments confirm that this set of sequences is effective in mice cell line.
Keywords
CRISPR-Ca9, RNA, DNA sequences
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[3] Ran, F.A., et al., Genome engineering using the CRISPR-Cas9 system. Nature Protocols, 2013. 8(11): p. 2281-2308.
[4] Porteus, M.H. and D. Carroll, Gene targeting using zinc finger nucleases. Nature Biotechnology, 2005. 23(8): p. 967-973.
[5] Sommer, D., et al., TALEN-mediated genome engineering to generate targeted mice. Chromosome Research, 2015. 23(1): p. 43-55.
[6] CRISPR-CAS9, TALENS and ZFNS - the battle in gene editing. Available from: https://www.ptglab.com/news/blog/crispr-cas9-talens-and-zfns-the-battle-in-gene-editing/
[7] Press release: The Nobel Prize in Chemistry 2020. Nobel Foundation. Retrieved 7 October 2020. Available from: https://www.nobelprize.org/prizes/chemistry/2020/press-release/
Copyright © 2021 Xijing Liao, Xueyan Yang
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