re-writing genomes: discoveries to applications: jennifer doudna
TRANSCRIPT
Re-writing Genomes: Discoveries to Applications
BASF Symposium June 23-24 2015Jennifer Doudna, UC Berkeley/HHMI
A cell’s DNA holds the key to its growth and development
What if a cell’s DNA could be edited just like the text of a document…
A revolutionary genome engineering technology began with…
curiosity about how bacteria fight the flu
CRISPRs: Hallmarks of acquired immunity in bacteria
Clusters of Regularly Interspaced Short Palindromic Repeats (CRISPRs)
Bolotin et al. 2005Mojica et al. 2005Pourcel et al. 2005
Bacterial/archaealchromosome
CRISPR locus
Jillian Banfield, UC Berkeley
How genetic information is stored and transferred
Sometimes RNA molecules function without encoding proteins
Three steps to acquire immunity in bacteria
Reviews: Horvath & Barrangou (2010) Science; Wiedenheft et al. (2012) Nature
Viral DNA
Emmanuelle CharpentierJennifer DoudnaMartin JinekKrzysztof ChylinskiInes Fonfara(Michael Hauer)
CRISPR-Cas9 Team
A programmable protein that cleaves DNA: Cas9
The Aha! Moment…
Cas9 protein
DNA
RNA
Cas9 and its guide RNA act like a molecular scalpel to cut DNA
Cas9 protein
DNA
RNA
Cas9 and its guide RNA act like a molecular scalpel to cut DNA
Programming Cas9 with single-guide RNAs(sgRNAs)
Jinek et al. Science 337, 816 (2012)
Programmed Cas9 cleaves DNA at specified sites
Jinek et al. Science 337, 816 (2012)
Genome editing begins with dsDNAcleavage
Cas9:sgRNA
June 2012
Exponential growth of genome engineering!
Courtesy of Jan Witkowski, Banbury Center
Prof. Russell Vance, UC Berkeley
An example of germline editing
Prof. Russell Vance, UC Berkeley
Inactivating a black coat color gene
How does Cas9 find a target site in a genome?
DNA target search and recognition by Cas9:gRNA
Sternberg et al. (2014) Nature
RNA-induced conversion of Cas9 into an active conformation for DNA surveillance
Jinek, Jiang, Taylor, Sternberg et al. (2014) Science
RNA-induced conversion of Cas9 into an active conformation for DNA surveillance
Jinek, Jiang, Taylor, Sternberg et al. (2014) Science
2.9 Å resolution
Jiang et al. (2015) Science, in press
Tracking Cas9 in Living Cells
Diffusion/search dynamics
Spencer Knight, James Liu, Robert Tjian
What should we do now that genomes can be edited quickly and easily?
New York Times:
MIT Technology Review:
Forbes:
Sy ReddingEric Greene, Columbia University
Eva Nogales, UC Berkeley
Alan Engelman, Harvard Medical School
Krys ChylinskiEmmanuelle Charpentier, Braunschweig
Acknowledgments