Introduction:
President Gast welcomed attendees to the 33rd Schrodinger Lecture, an annual event celebrating the brilliance and lasting influence of Professor Erwin Schrodinger’s work. The virtual format allowed for a larger audience than ever before. The lecture honors Professor Schrodinger’s memory and provides a platform for exciting new discoveries by world-renowned researchers.

Imperial’s Connection to Schrodinger:
Imperial has a special connection to Schrodinger, having hosted a celebration of his birth centenary 34 years ago. Professor Terry Rudolph, Schrodinger’s grandson and Imperial’s professor of quantum physics, was present along with other family members. The Schrodinger Scholarships attract outstanding PhD students in natural sciences, and philanthropy plays a vital role in supporting these talented scholars.

Professor Jennifer Doudna: Speaker and Nobel Laureate:
The speaker for the 33rd Schrodinger Lecture was Professor Jennifer Doudna, a world leader in biochemistry and genetics, Nobel laureate, and awe-inspiring speaker. There is a direct link between Professor Doudna and Professor Schrodinger, as Schrodinger’s book “What is Life?” inspired James Watson and Francis Crick’s research into the double helix, which in turn inspired Professor Doudna’s interest in genetics.

Schrodinger’s Contributions:
Professor Schrodinger is best known for developing quantum theory, earning him the 1933 Nobel Prize in physics. His book “What is Life?” explored the idea of genetic information encoded in the configuration of chemical bonds, inspiring research into the double helix.

Inspiration and Legacy:
Professor Doudna’s passion for science was sparked by her father’s gift of the book “The Double Helix” by James Watson. The Schrodinger Lecture serves as a tribute to Professor Schrodinger’s brilliance and lasting influence on scientific research.

What is CRISPR?:
CRISPR is a revolutionary gene-editing technology that enables scientists to alter DNA, the code of life. This technology is based on a bacterial immune system called CRISPR-Cas9, which helps bacteria defend against viruses.

Genome Editing with CRISPR:
CRISPR-Cas9 can be programmed to target specific DNA sequences and make precise changes to the genome. This technology has opened up new possibilities for treating genetic diseases, improving crop yields, and even creating new materials.

Clinical Applications of CRISPR:
CRISPR is already being used in clinical trials to treat a variety of genetic diseases. One promising area of research is the use of CRISPR to correct genetic mutations that cause sickle cell disease. CRISPR is also being studied as a potential treatment for cancer and HIV.

Advancements in CRISPR Technology:
Ongoing research is focused on advancing CRISPR technology for clinical use. This includes developing more efficient and accurate ways of delivering CRISPR to specific cells. Researchers are also working to reduce the risk of off-target effects, where CRISPR makes unintended changes to the genome.

Conclusion:
CRISPR is a powerful gene-editing technology with a wide range of potential applications in biology and medicine. Clinical trials are already underway, and CRISPR holds great promise for treating a variety of genetic diseases. Researchers are actively working to improve the technology and make it safer and more effective for use in patients.

What is CRISPR?:
CRISPR is a groundbreaking technology that enables programmable genome editing, allowing precise changes to DNA sequences in cells.

Mechanism of CRISPR:
CRISPR works by utilizing a protein, CRISPR-Cas9, which recognizes and binds to specific DNA sequences. Once bound, CRISPR-Cas9 cuts the DNA at the targeted site, enabling cells to make precise changes to the DNA at the break site.

Origins of CRISPR:
CRISPR originated from the study of how bacteria fight viral infections. Bacteria possess CRISPR systems that allow them to acquire and store pieces of viral DNA, enabling them to recognize and destroy future viral infections.

CRISPR in Eukaryotic Cells:
In eukaryotic cells, double-stranded breaks in DNA can be repaired through various mechanisms. This repair process can introduce changes to the DNA sequence or integrate new genetic material at the break site.

Harnessing CRISPR for Genome Editing:
Understanding the DNA cleavage mechanism of CRISPR led to the realization that it could be harnessed as a tool for genome editing in other cell types. CRISPR-Cas9, guided by a specific RNA sequence, can be directed to target and cut DNA at desired locations in the genome.

Video Illustration:
A video animation is used to demonstrate the CRISPR-Cas9 system targeting and cutting DNA in a eukaryotic cell.

CRISPR Basics:
CRISPR-Cas9 is a revolutionary technology that enables targeted editing of DNA sequences. It utilizes a protein called Cas9, which forms an RNA-DNA helix at the match site and cuts both DNA strands. The double-stranded break is then repaired by cellular repair proteins, leading to the actual editing.

CRISPR-Cas9 Simplification:
Jennifer Doudna and Emmanuelle Charpentier’s research simplified the CRISPR system, making it easier to use. Chris Chylinski and Martin Jinek demonstrated that the system could be programmed to cut any desired DNA sequence.

CRISPR’s Potential:
CRISPR can induce targeted changes in the genome of virtually any cell type, opening up many opportunities for genome editing applications.

Applications of CRISPR:
CRISPR has various applications, including: Gene therapy: Treating genetic diseases by correcting faulty genes. Cancer immunotherapy: Enhancing immune cell activity against cancer cells. Crop improvement: Modifying crops for improved yield, resilience, and nutritional value. Basic research: Investigating gene function and regulation.

Precision Editing:
CRISPR has emerged as a powerful tool for manipulating genomic information, addressing the need for precise editing capabilities in various organisms, including butterflies and mammals. This technology has wide-ranging applications in fundamental research, medicine, and agriculture. In clinical medicine, CRISPR is being used as a tool of discovery and as an actual therapy for genetic disorders like sickle cell disease.

Clinical Application:
Sickle cell disease is caused by a single base pair mutation in the gene encoding human beta-globin protein, leading to abnormal hemoglobin and health complications. CRISPR has been successfully used to correct the disease-causing mutation in cells derived from sickle cell patients in the laboratory and is now showing promising results in clinical trials. The first patient to receive CRISPR treatment in the United States, Victoria Gray, has demonstrated the effectiveness of this therapy.

Germline Editing:
CRISPR raises ethical concerns regarding its potential use in the human germline, where heritable changes can be passed on to future generations. Discussions are ongoing to determine when and how CRISPR can be ethically applied in the germline, considering the possibility of treating certain diseases in the future.

Advancements in CRISPR Technology:
CRISPR and genome editing are rapidly advancing, with ongoing research to improve the precision and efficiency of editing techniques. Scientists are exploring methods to ensure the precision of editing, address challenges like off-target effects, and expand the range of genetic changes that can be made.

Future of CRISPR in Clinical Medicine:
CRISPR is poised to revolutionize clinical medicine by providing a standard of care for various genetic diseases. The technology holds immense potential for treating genetic disorders, improving agricultural crops, and addressing climate change challenges. CRISPR’s platform nature allows for continuous advancements and the development of innovative applications in the future.

A-Base Editing: A Powerful Tool for Precise DNA Modifications:
A-Base Editing (ABE) is an innovative technique that combines the power of CRISPR-Cas9 with specific biochemical domains to achieve targeted DNA base pair changes. This technology enables the conversion of an adenosine (A) to an inosine (I) and subsequently to a guanine (G) within the DNA sequence.

Targeting Single Nucleotide Polymorphisms Associated with Diseases:
ABE addresses approximately half of the known human single nucleotide polymorphisms (SNPs) associated with diseases. These SNPs, which are single-base pair variations, can potentially be corrected by altering AT base pairs to GC base pairs.

TADA’s Role in ABE:
A crucial component of ABE is the TADA protein, derived from bacteria. TADA is a tRNA editing enzyme that has undergone directed evolution to recognize and modify single-stranded DNA. This adaptation allows TADA to function in conjunction with the CRISPR-Cas9 system.

CRISPR-Cas9 Facilitates Single-Stranded DNA Access:
The CRISPR-Cas9 system plays a critical role in ABE by facilitating access to single-stranded DNA. The natural mechanism of CRISPR-Cas9 involves melting open the DNA duplex, exposing the single-stranded region where TADA can perform its editing function.

Challenges in Optimizing ABE:
The development of ABE faced several challenges, including off-target RNA editing and the need to enhance the editing activity of the proteins. Iterative rounds of laboratory evolution and testing were conducted to address these issues and improve the overall performance of ABE.

ABE8E’s Remarkable Efficiency:
Among the various versions of the CRISPR-Cas9 A-base editor tested, ABE8E, developed by the Lou Lab, exhibited exceptional efficiency. This enzyme demonstrated significantly faster production of edited products compared to other variants.

Basic Function of TAD-A Base Editor:
TAD-A base editor is a remarkably fast enzyme for editing DNA sequences. It can carry out multiple editing reactions per enzyme molecule, which is unique compared to other base editors.

Chemical Trapping of TAD-A Enzyme:
A collaboration with Peter Beal’s lab at UC Davis utilized a chemical trapping strategy to capture TAD-A in an intermediate step of its editing reaction. An analog called 8-azonebularin was used to stop the reaction and create a stable intermediate analog.

Structural Analysis:
This trapping strategy allowed researchers to obtain a structure of the TAD-A base editor using cryo-electron microscopy. The structure revealed that the editor sits on the DNA at a site made accessible by CRISPR-Cas9. Mutations introduced during in vitro selection stabilized the structure of the DNA required for editing.

Significance:
Understanding the structure of the TAD-A base editor provides insights into its fast and efficient editing activity. This knowledge can aid in designing strategies to enhance editing specificity and prevent unwanted off-target edits.

Cas5: A Tiny CRISPR Enzyme:
CRISPR-Cas9 is not the only CRISPR-Cas enzyme. Cas5 is a tiny CRISPR enzyme encoded in phage. Cas5 has its own CRISPR array in the phage genome that creates RNAs that guide its activity.

Features of Cas5:
Cas5 is very tiny and compact. Cas5 has the smallest molecular weight of all known CRISPR-Cas enzymes. Cas5 may be easier to understand its molecular functions and have chemical properties that lend themselves to delivery into different cells.

Experiment Testing Cas5 Activity:
Cas5 was introduced into cultured human embryonic kidney cells. A piece of DNA encoding Cas5 protein with appropriate guide RNAs was used. The experiment targeted a gene encoding a green fluorescent protein, allowing easy identification of edited cells.

Structural Studies of Cas5:
Cas5 has a tight interaction with its guide RNA. Cas5 can unwind duplex DNA. Ongoing research aims to understand Cas5’s biochemical functions and potential applications.

Delivery of CRISPR-Cas Editing Enzymes:
The question of how CRISPR-Cas editing enzymes can be delivered into specific cell types is being addressed.

Challenges in CRISPR-Mediated T Cell Therapy:
The primary challenge in CRISPR-based T cell therapy lies in efficiently delivering pre-assembled Cas guide RNA complexes into target cells.

Current Approach:
Researchers are exploring the creation of virus-like particles (VLPs) to serve as delivery vehicles for pre-assembled Cas guide RNA complexes.

VLP Design and Strategy:
The VLPs are designed to target genes relevant to T cell engineering. They lack infectious genes but retain genes necessary for capsid formation. The VLPs also contain proteins projecting from the surface that facilitate recognition and entry into specific cell types.

Encoding and Packaging of CRISPR Components:
The VLPs encode proteins required for capsid formation along with a fusion protein called Gag Cas9. Gag Cas9 enables the expression of Cas9, allowing for packaging into capsids. Cleavage from the membrane releases the assembled protein guide RNA inside the capsid.

Co-Delivery of Donor DNA:
In some cases, the VLPs can co-deliver a piece of donor DNA. This donor DNA provides information for DNA repair, enabling the integration of new genetic information following Cas9-mediated cutting.

Applications:
This approach allows for the creation of chimeric antigen receptors (CARs) in T cells, enhancing their ability to recognize and attack cancer cells.

CRISPR-Cas9 Technology for Cancer Immunotherapy:
Researchers have employed CRISPR-Cas9 to modify T cells, a crucial component of the immune system, for cancer immunotherapy. These modified T cells can target and eliminate cancer cells more effectively, offering a promising approach for cancer treatment. Techniques have been developed to edit specific types of T cells, allowing precise targeting of cancer cells.

CRISPR-Cas9 for Gene Editing in the Brain:
Modified forms of CRISPR-Cas9 have been engineered to penetrate the blood-brain barrier and enable gene editing in the brain. This technology holds potential for treating neurological disorders such as Huntington’s disease and glioblastoma.

CRISPR for Diagnostics:
CRISPR-Cas systems can recognize specific RNA sequences and activate a reporter enzyme to generate a fluorescent signal. This mechanism forms the basis of CRISPR diagnostics, which provide rapid and sensitive detection of RNA sequences, including pathogens like SARS-CoV-2.

Future Directions:
Understanding the molecular mechanisms of CRISPR enzymes is vital for developing more efficient and accurate tools for genome editing and beyond. The discovery of new CRISPR proteins, such as compact editors, expands the options for clinically useful tools. Research is ongoing to enable in situ delivery of CRISPR enzymes in patients, making the technology more accessible and affordable.

Ethical Considerations:
Promoting public confidence in genome editing requires scientific engagement, transparency, and accurate communication about the technology’s capabilities and limitations. Addressing ethical concerns is crucial, especially in areas such as germline editing and human enhancement.

Challenges Faced by Women in Science:
Women in science often face self-doubt and may require more confidence-building support, particularly early in their careers. Mentorship from encouraging and supportive individuals, both male and female, plays a significant role in overcoming these challenges.

CRISPR in Agriculture:
CRISPR technology has the potential to revolutionize agriculture and address global food insecurity. Its applications include enhancing crop resilience, resistance to pests and diseases, and improving nutritional content. CRISPR offers opportunities for developing new and improved varieties of crops, contributing to sustainable food production.

CRISPR Technology’s Practical Impacts:
CRISPR-modified plants with desirable traits, such as mushrooms that resist browning, are already entering the market. Future applications include crops with increased yields, drought and pest resistance, and enhanced nutritional value, benefiting global food production and sustainability.

Ethical Considerations for High School Students:
Students should critically assess CRISPR technology’s ethical implications through the lens of equity and health. The technology’s transformative potential demands responsible stewardship and consideration of its impact on future generations.

Affordability and Accessibility:
Ensuring the affordability and accessibility of CRISPR-based treatments is crucial for equitable access to life-saving therapies. Collaborative efforts are needed to make the technology accessible to underserved populations and developing countries.

Public Access to CRISPR:
Broad access to CRISPR in non-traditional scientific environments can empower individuals to explore and utilize the technology. However, responsible use and comprehensive safety guidelines are essential to mitigate potential risks associated with DIY genetic engineering.

Safeguards for Human Genome Editing:
Transparency, open discussion, and global collaboration are vital for establishing safeguards and ethical standards for human genome editing. Individual responsibility and the scientific community’s collective stance on responsible deployment are crucial in shaping the future of the technology.

Timeline of CRISPR’s Impact:
CRISPR’s initial impact may be seen in agriculture, with genetically modified fruits and vegetables entering markets. Cancer immunotherapy and personalized medicine are other areas where CRISPR is expected to make a significant impact. Delivery strategies for CRISPR-Cas to specific cell types need further development for effective treatment of diseases like muscular dystrophy and cystic fibrosis.

Personalized Medicine:
Predicting and preventing diseases through genetic analysis and targeted gene editing is a potential future application of CRISPR. However, extensive research and safety considerations are necessary before such interventions can be widely implemented.

Medical Regulation:
Existing regulatory frameworks for molecular cloning provide a foundation for CRISPR regulation. Unique challenges in personalized medicine applications may require additional regulatory measures to ensure safety and efficacy.

Challenges in CRISPR Clinical Trials:
Ensuring the safety and effectiveness of CRISPR technology while maintaining a practical and efficient clinical trial process is a current challenge.

Book Recommendations for Genetics and Genetic Engineering:
Books by Lewis Thomas, Harold Morowitz, and Sid Mukherjee provide insightful explanations of genetics. The Eighth Day of Creation is a notable book that presents genetics in an engaging manner.

Multimedia Resources for Younger Audiences:
Khan Academy offers effective videos that explain the fundamentals of genetics and CRISPR.

Entrepreneurship in Teaching:
The speaker’s thoughts on embedding entrepreneurship in teaching are still evolving.

Jennifer Doudna’s Background and Shift in Perspective:
Doudna’s early education emphasized traditional academic pursuits, with a focus on understanding the fundamentals of nature. Later, she recognized that certain scientific advancements reach a point where the next steps are better suited for non-academic settings due to factors like cost, practicality, and educational limitations.

Importance of Distinguishing Academic and Commercial Endeavors:
Doudna stresses the significance of teaching students to differentiate between scientific projects best suited for academic environments and those more appropriate for focused commercial endeavors. This distinction is crucial as students face increasing opportunities to pursue careers in companies after scientific training.

Entrepreneurship Opportunities for Scientists:
In contrast to the limited opportunities during her PhD, Doudna’s students now have various options to engage in entrepreneurship, allowing them to pursue scientific careers outside academia.

Potential Breakthrough to Accelerate CRISPR’s Impact:
Doudna identifies efficient delivery of CRISPR-Cas into desired cell types as a key breakthrough that could revolutionize the real-world value and impact of CRISPR technology. Effective delivery would have a profound impact in clinical medicine and agriculture.

Closing Remarks and Gratitude:
Doudna expresses her gratitude for the opportunity to lecture and answer questions, despite the virtual nature of the event. She appreciates the invitation and acknowledges the challenges of conducting the talk remotely.