2:15 – 2:45, 6/13/2014, Jennifer Doudna “The biology of CRISPRs: from genome defense to genetic engineering”
Reporter: Aviva Lev-Ari, PhD, RN
LIVE from Kresge Auditorium @MIT
About CRISPR “this technology will revolutionize biology in the same way PCR did,” Rudolf Jaenisch introducing Jennifer Doudna
Top CRISPR Related Publications
http://blog.appliedstemcell.com/top-crispr-related-publications/
What is CRISPR? Why are Cas9-CRISPR services so popular?
http://blog.appliedstemcell.com/what-is-crispr-why-are-cas9-crispr-services-so-popular/
Custom Rat Model Generation Service Using CRISPR/Cas9
http://www.appliedstemcell.com/services/animal-models/
Capturing key concepts of Prof. Jennifer Doudna’s Lecture @ KI Symposium:
- acquired immunity in bacteria
- three steps:
- adaptation
- biogenesis
- interference
- type I: e-coli – cascade recognize foreign DNA
- type II: S. pyogenes (human) gene for DNA targeting
- Cas9 is a dual RNA guided dsDNA endonuclease
- Programming Cas9 with single-guided RNAs (sgRNAs)
- Cleaving DNA at specific sites
- Programmable DNA targeting by Cas9:gRNA: Specific genome editing: transcriptional control
- Cas9 – protein – how it works? guide RNA and clivage activity
- Cas9:gRNA to phage Gamma DNA: PAM binding activates Cas9 nuclease domain
- Mechanism of DNA interrogation: high-affinity product binding, DNA duplex guided by RNA
- Binding – gRNA:DNA heteroduplex formation is directional, RNA-induced conversion of Cas9 into an active confirmation for DNA surveillance
- Cutting
- DNA target search and recognition by Cas9:gDNA
- Why Cas9 targets DNA and not RNA?
- Programmed RNA cleavage using Cas9:gRNA: dsDNA, ssRNA, PAMmer (19-nt)
Conclusions
Cas9:RNA specifically bind PAM sites
PAM Recognition
2:15 – 2:45, 6/13/2014, Jennifer Doudna “The biology of CRISPRs: from genome defense to genetic engineering”
@MIT – Summer Symposium 2014: RNA Biology, Cancer and Therapeutic Implications, June 13, 2014 8:30AM – 4:30PM, Kresge Auditorium @MIT
http://ki.mit.edu/news/symposium
REALTIME event coverage for the Scientific Media by Dr. A. Lev-Ari
in Open Access Scientific Journal of Leaders in Pharmaceutical Business Intelligence (LPBI)
http://pharmaceuticalintelligence.com
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Jennifer Doudna, PhD
Professor of Molecular and Cell Biology and Chemistry, University of California, Berkeley
Investigator, Howard Hughes Medical Institute
Jennifer A. Doudna, PhD, Professor of Molecular and Cell Biology and Chemistry at the University of California, Berkeley and Howard Hughes Medical Institute investigator, has devoted her scientific career to revealing the secret life of RNA. Using the approaches of structural biology and biochemistry, Doudna’s work deciphering the molecular structure of RNA enzymes and other functional RNAs has shown how these seemingly simple molecules can carry out complex functions and can work together with proteins to control the information content of a cell.
Doudna grew up amidst the natural wonders of Hawaii, where she experienced volcanic eruptions, explored remote beaches and honed her body-surfing skills while living in the small town of Hilo on the Big Island. Doudna earned a BA in biochemistry at Pomona College in 1985, where she worked with outstanding chemists Sharon Panasenko and Fred Grieman, and enjoyed the mentorship of many other great professors. She then worked with Jack Szostak at Harvard, completing her PhD in 1989 on the develpoment of a self-replicating RNA based on the activity of a group I self-splicing intron. This work showed how RNA could function as both a template and a catalyst for generating copies of itself, a key propoerty of life. As a Lucille Markey postdoctoral associate with Tom Cech at University of Colorado at Boulder, Doudna began crystallizing catalytic RNA molecules with a goal of determining their three-dimensioanl structures and hence inlocking the key to their biochemical activities. She continued this work as a faculty member at Yale University, where she became a professor in 1994 in the Department of Molecular Biophysics and Biochemistry. In two landmark studies early in her career, Doudna and colleagues solved the crystal structures of two large RNAs – the P4-P6 domain of the Tetrahymena thermophila group I intron ribozyme and the hepatitis delta virus ribozyme. By determining their molecular structures, her work advanced the understanding of RNA’s ability to function as a catalyst in biological systems. In 2002, Doudna moved to the University of California at Berkeley, where her lab began studying the function of small RNAs that control the use of a cell’s genetic information. This led to her work on bacterial immune systems that employ RNA molecules derived from viruses to target and destroy foreign DNA. In collaboration with the lab of Emmanuelle Charpentier, Doudna and postdoctoral associate Martin Jinek discovered the function of an RNA-guided enzyme in the bacterial immune pathway, Cas9, whose ability to cut double-stranded DNA can be programmed by changing the guide RNA sequence. They recognized that such an activity could be employed as a molecular tool for precision genome engineering in various kinds of cells, a discovery that has triggered a revolution in the fields of molecular genetics and genomics.
Doudna’s work has been honored by numerous awards. She received the National Academy of Sciences Award for Initiatives in Research in 1999, and the Alan T. Waterman Award from the NSF in 2000. In 2001 she received the Eli Lilley Award in Biological Chemistry from the American Chemical Society, and in 2013 she was the recipient of the Mildred Cohn Award from ASBMB and the Hans Neurath Award from the Protein Society. She has been a Howard Hughes Medical Institute investigator since 1997 and a member of the National Academy of Sciences since 2002. She was named to the American Academy of Arts and Sciences in 2003 and elected to the Institute of Medicine in 2010. In 2014 she received the Lurie Prize from the Foundation for the NIH.
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