Jennifer A. Doudna Adam Grosvirt-Dramen Hochbaum Lab Group Meeting 10/22/2020

Super scientist: Jennifer A. Doudna Adam Grosvirt-Dramen Hochbaum Lab Group Meeting 10/22/2020

https://www.emmanuelle-charpentier-lab.org/ Who is a scientist (living or dead) that you admire? In what field did they work?

• Dr. Jennifer A. Doudna • Biochemistry professor at UC Berkeley working on gene editing using CRISPR technology • Chemistry Nobel Laureate 2020 Summarize their career history - where did they study, what fields, any non-academic pursuits? How did they get to the point in their career when they made significant impact on their field?

B.A. Chemistry Ph.D. Biochemistry, 1989 Lucille P. Markey Scholar in Henry Ford II Professor of 1985 Post-Doc, 1989-1991 Biomedical Science, 1991-1994 Molecular Biophysics and Dr. Sharon Panasenko Dr. Jack W. Szostack Dr. Thomas R. Cech Biochemistry, 1994-2002 (Nobel 2009) (Nobel 1989)

Professor of Biochemistry and Molecular Biology at UC Berkeley and Faculty Scientist at LBNL in the Physical Biosciences Division 2003-Present https://doudnalab.org/people/ Summarize their career history - where did they study, what fields, any non-academic pursuits? How did they get to the point in their career when they made significant impact on their field?

COMMON RESEARCH THEME

• Used CRISPR Cas-9 to edit the genome of bacteria using a guide RNA sequence to target specific DNA sequences • Charpentier found the system in bacteria • Doudna used this system to edit bacterial genomes • Edit the genome  Endless possibilities • cure diseases • fight off viruses • plant foods that are resistant to the elements and pests

https://www.nobelprize.org/prizes/chemistry/2020/advanced-information/ Background: What was/is the scientific context of their work? What was the understanding in their field at the time of their work? Put into context the problem or advancement they made. • Bacteria have genetic sequences that act as a history of viral infections • CRISPR = clustered regularly interspaced short palindromic repeats • CRISPR + Cas proteins act as bacterial immune system to break down invasive plasmids or viral DNA • Nobel Prize is for using targeted sequences and expressing CRISPR and Cas in cells to change genetic sequences What was the key conceptual insight(s) that enabled the scientist's discovery(ies), and how are they different from what others were thinking?

Protospacers in CRISPR array is the library of immunity

Artificially introducing new repeats or taking them away can effect immunities and defense against viruses and pathogens

Removing protospacers removed the immunity from history What were key papers that led to the advancements the scientist is known for? What experimental design/setups produced key results and how? How were these data interpreted and how do they support their discoveries?

• Paper for which Doudna and Charpentier won the Nobel Prize • tracrRNA combined with crRNA triggers Cas9 to break down foreign DNA • Cas9 is an endonuclease that breaks apart internal bonds in DNA rather than just at the 3’ or 5’ end • Just overexpressing Cas9 in bacteria doesn’t break down the foreign DNA • Site specific cleavage of intruding DNA that pairs with crRNA • Single RNA molecule with tracrRNA and crRNA features can be used to guide Cas9 proteins to break apart DNA in specified locations https://science.sciencemag.org/content/337/6096/816.long What were key papers that led to the advancements the scientist is known for? What experimental design/setups produced key results and how? How were these data interpreted and how do they support their discoveries?

• Paper for which Doudna and Charpentier won the Nobel Prize • tracrRNA combined with crRNA triggers Cas9 to break down foreign DNA • Cas9 is an endonuclease that breaks apart internal bonds in DNA rather than just at the 3’ or 5’ end • Just overexpressing Cas9 in bacteria doesn’t break down the foreign DNA • Site specific cleavage of intruding DNA that pairs with crRNA • Single RNA molecule with tracrRNA and crRNA features can be used to guide Cas9 proteins to break apart DNA in specified locations Activates in order to bind to DNA https://science.sciencemag.org/content/337/6096/816.long What were key papers that led to the advancements the scientist is known for? What experimental design/setups produced key results and how? How were these data interpreted and how do they support their discoveries?

• 5 target sequences in a dsDNA sequence encoding for GFP • Cas cleaves at designated site along the dsDNA • Each site has its own corresponding chimeric RNA that targets the site to be cleaved • Control sample (right column) was not cleaved • Entire 11100 bp dsDNA sequence • Each chimeric RNA plus Cas-9 protein complex cut at the precise point that the target RNA codes for

https://science.sciencemag.org/content/337/6096/816.long What are the current (and future) directions/applications made possible by the scientist's discoveries?

https://www.royalsociety.org.nz/what-we-do/our-expert-advice/all-expert-advice-papers/gene-editing-technologies/current-gene-editing-uses/ Who were key collaborators and researchers that enabled these discoveries?

• Emmanuelle Charpentier equally shares the Nobel Prize in Chemistry 2020 with Jennifer Doudna • Scientific and Managing Director at the Max Planck Unit for the Science of Pathogens in Berlin, Germany • Honorary Professor, Institute of Biology at Humbolt University in Berlin Germany • World-leading expert in regulatory mechanisms underlying processes of infection and immunity in bacteria • Discoveries in molecular pathways of antibiotic resistance and bacteria virulence • Charpentier discovered the CRISPR immune response in bacteria and then teamed up with Doudna to further develop the CRISPR-Cas protein genome editing process Include an additional fun fact about the scientist.

• We share the same birthday: Feb 19 • Grew up in Hawaii to parents who are professors in humanities and social science • The lush plants and her chemistry class in high school gave her an appreciation for nature that continues into her current research • Spent a summer studying mushrooms with at the university of Hawaii at Hilo with Don Hemmes (family friend) • Leadership from her female undergrad advisor helped inspired her to pursue a science career • “Mentors are critical” – Interview with UC Berkeley’s California Magazine