Abstract
The CRISPR-Cas9 technology is a powerful tool that enables site-specific genome modification (gene editing) and is increasingly used in research to generate gene knockout or knock-in in a variety of cells and organisms. This chapter provides a brief overview of this technology and describes a general methodology applicable to human airway biology research.
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References
Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S et al (2007) CRISPR provides acquired resistance against viruses in prokaryotes. Science 315:1709–1712
Brouns SJ, Jore MM, Lundgren M, Westra ER, Slijkhuis RJ, Snijders AP et al (2008) Small CRISPR RNAs guide antiviral defense in prokaryotes. Science 32:960–964
Marraffini LA, Sontheimer EJ (2008) CRISPR interference limits horizontal gene transfer in staphylococci by targeting DNA. Science 322:1843–1845
Ishino Y, Shinagawa H, Makino K, Amemura M, Nakata A (1987) Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product. J Bacteriol 169:5429–5433
Mojica FJ, DÃez-Villaseñor C, Soria E, Juez G (2000) Biological significance of a family of regularly spaced repeats in the genomes of Archaea, Bacteria and mitochondria. Mol Microbiol 36:244–246
Bolotin A, Quinquis B, Sorokin A, Ehrlich SD (2005) Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. Microbiology 151:2551–2561
Pourcel C, Salvignol G, Vergnaud G (2005) CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA, and provide additional tools for evolutionary studies. Microbiology 151:653–663
Marraffini LA (2015) CRISPR-Cas immunity in prokaryotes. Nature 526:55–61
Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE et al (2013) RNA-guided human genome engineering via Cas9. Science 339:823–826
Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F (2013) Genome engineering using the CRISPR-Cas9 system. Nat Protoc 8:2281–2308
Mojica FJ, DÃez-Villaseñor C, GarcÃa-MartÃnez J, Almendros C (2009) Short motif sequences determine the targets of the prokaryotic CRISPR defense system. Microbiology 155:733–740
Bauer DE, Canver MC, Orkin SH (2015) Generation of genomic deletions in mammalian cell lines via CRISPR/Cas9. J Vis Exp 95:e52118
Acknowledgments
This work was supported by the following grants from NIH: 1U19AI125357, R01HL122321, R01AI106287, and R01HL125128.
The authors wish to thank Max Seibold, Jamie Everman, and Ari Stoner (Dr. Max Seibold’s Lab, National Jewish Health, Denver) for technical advice and support.
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Dakhama, A., Chu, H.W. (2018). The Use of CRISPR-Cas9 Technology to Reveal Important Aspects of Human Airway Biology. In: Reinhardt, R. (eds) Type 2 Immunity. Methods in Molecular Biology, vol 1799. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-7896-0_27
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DOI: https://doi.org/10.1007/978-1-4939-7896-0_27
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