This series of short presentations on gene editing is brought to you by Dr Adam West, College of Medical Veterinary and Life Sciences at the University of Glasgow, Scotland.
www.gla.ac.uk/people/adamwest
This presentation describes the use of S.pyogenes CRISPR/Cas9 system for genome editing, including:
2:50 How to deliver to cells
10:12 How guide RNAs are expressed from plasmids
11:26 How to assay for CRISPR-directed mutagenesis
This is part of a series ( • Genome Editing Explained ) that also covers
Zinc finger nucleases (ZFNs)
TALE nucleases (TALENs)
CRISPR adaptive immune systems
CRISPR specificity
Precision editing
Homology directed repair
Prime editing
This presentation is primarily aimed at university students, researchers, clinicians and journalists interested in fields related to molecular biology and genetics. It is for education purposes only.
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More accessible presentations for a wide audience can be found in the “Explained Simply” section of this channel in the near future.
Artwork is © Adam West with the exception of images and data taken from publications referenced in these slides. References to reviews and journal articles are denoted by circled numbers at the bottom right of the slides.
Links to these numbered articles are listed below. We recommend these as your best way to begin further reading on this subject.
Publication List
17. Mali P, Esvelt KM, Church GM.
Cas9 as a versatile tool for engineering biology.
Nat Methods. 2013 Oct;10(10):957-63.
doi.org/10.1038/nmeth.2649
19. Garside EL, MacMillan AM.
Cas9 in close-up.
Nat Biotechnol. 2014 Apr;32(4):338-40.
doi.org/10.1038/nbt.2872
20. Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O.
Crystal structure of Cas9 in complex with guide RNA and target DNA.
Cell. 2014 Feb 27;156(5):935-49.
doi.org/10.1016/j.cell.2014.0...
21. Hsu PD, Lander ES, Zhang F.
Development and applications of CRISPR-Cas9 for genome engineering.
Cell. 2014 Jun 5;157(6):1262-78.
doi.org/10.1016/j.cell.2014.0...
23. Liang X, Potter J, Kumar S, Zou Y, Quintanilla R, Sridharan M, Carte J, Chen W, Roark N, Ranganathan S, Ravinder N, Chesnut JD.
Rapid and highly efficient mammalian cell engineering via Cas9 protein transfection.
J Biotechnol. 2015 Aug 20;208:44-53.
doi.org/10.1016/j.jbiotec.201...
24. Brinkman EK, Chen T, Amendola M, van Steensel B.
Easy quantitative assessment of genome editing by sequence trace decomposition.
Nucleic Acids Res. 2014 Dec 16;42(22):e168.
doi.org/10.1093/nar/gku936
Music credits
“Go, Icarus! Go!” by The Whole Other
“New Year” by Bad Snacks
(RU-vid Audio Library)
17 фев 2020
