Background of the Bampton Lectures:
The Bampton Lectures were established by Ada Bampton Tremaine in the early 20th century and are given periodically in various fields, including religion, science, art, and medicine.

Introduction of Professor Jennifer Doudna:
Professor Jennifer Doudna is a renowned chemist and molecular and cell biologist at Berkeley. She is particularly known for her 2012 article in Science, co-authored with Emmanuel Charpentier, which presented a simple method for editing DNA using an RNA-guided protein found in bacteria.

Significance of CRISPR-Cas9 Discovery:
CRISPR-Cas9, the procedure described in the 2012 article, was recognized as a groundbreaking advancement in genome editing. It has led to numerous awards and research exploring the possibilities of genome editing in various applications.

Previous Lecture:
Professor Doudna’s previous lecture at the Medical Center focused on the science of CRISPR and recent research in the field, particularly in her lab.

Ethical and Social Implications:
Professor Doudna has been actively engaged in discussions about the ethical and social implications of gene editing.

Upcoming Talk and Response:
Professor Doudna’s lecture today is titled “CRISPR Biology and Technology, the Future of Genome Editing.” Following her talk, Professor Hank Greeley, a law professor at Stanford specializing in ethical, legal, and social issues in biosciences, will provide a response. After the response and a brief dialogue with Professor Doudna, the session will be opened for questions from the audience.

Introduction:
Genome editing technology allows for precise and accurate changes to DNA in cells, opening up possibilities for disease correction and various biological manipulations.

Origins in Bacterial Research:
Curiosity-driven research on how bacteria fight viral infections led to the discovery of CRISPR, an adaptive immune system in bacteria. CRISPR involves acquiring viral DNA fragments and integrating them into the bacterial genome as a genetic vaccination card. When matching viral DNA is detected, CRISPR-associated proteins use RNA address labels to guide DNA destruction, protecting the cell from future infection.

Diversity of CRISPR Systems:
CRISPR systems vary in complexity, with some requiring multiple genes and others relying on a single protein, Cas9. Jennifer Doudna’s lab studied simpler CRISPR systems, focusing on the function of Cas9, a single enzyme providing RNA-guided protection.

Cas9: A Precise DNA Cleaver:
Cas9 is a protein directed by RNA to find and destroy viruses. It uses a 20-letter RNA sequence to recognize a matching sequence in DNA, unwinding the DNA double helix and generating a break. The specificity of Cas9’s cutting is dictated by the RNA guide, allowing for targeted DNA cleavage.

Single Guide RNA: A Technological Breakthrough:
Martin Yinek, a postdoc in Doudna’s lab, linked two RNA molecules in nature to form a single guide RNA. This allowed for the design of single guide RNAs with different address labels, enabling Cas9 to find and cut any desired DNA molecule. This discovery opened up the possibility of genome editing in eukaryotic cells, leading to a more profound and transformative technology.

Genome Editing Applications:
Double-stranded DNA breaks in eukaryotic cells can be repaired, resulting in either small disruptions or larger changes to the DNA sequence. This enables targeted gene correction, insertion, or deletion, with potential applications in medicine, agriculture, and basic research.

Introduction to Genome Editing:
Scientists have been studying DNA repair pathways, particularly double-strand break repair, to explore the possibility of altering genetic code at specific gene locations. The ability to make desired genetic changes with precision has significant implications for understanding gene functions and developing therapeutic approaches.

CRISPR-Cas9: A Revolutionary Tool:
The discovery of the CRISPR-Cas9 system, derived from bacterial defense mechanisms, revolutionized genome editing technology. CRISPR-Cas9 allows scientists to program a single protein (Cas9) to cut DNA at precise locations guided by a specific RNA molecule.

Mechanism of CRISPR-Cas9:
CRISPR-Cas9 acts like a molecular scissor, searching through DNA for a sequence that matches the guide RNA it carries. Upon finding a match, it cuts the DNA, creating a double-strand break. Cellular repair mechanisms then fix the break, providing an opportunity to introduce genetic changes at the cut site.

Applications of CRISPR-Cas9:
CRISPR-Cas9 has broad applications in various fields, including medicine, agriculture, and basic research. In medicine, CRISPR-Cas9 is being explored for gene therapy to correct genetic defects, develop new treatments for diseases like cancer and sickle cell anemia, and enhance immune system functions. In agriculture, CRISPR-Cas9 can be used to improve crop yields and resistance to pests and diseases. In basic research, CRISPR-Cas9 is a powerful tool for studying gene functions and unraveling the genetic basis of complex biological processes.

Potential Challenges and Ethical Considerations:
While CRISPR-Cas9 holds immense promise, it also raises ethical concerns, particularly regarding the potential unintended consequences of genetic modifications. The need for responsible and ethical guidelines to ensure the safe and responsible use of this technology is emphasized.

Introduction of CRISPR Technology and Its Rapid Advancement:
CRISPR technology has revolutionized the field of genome editing, leading to an exponential increase in scientific publications.

Mechanism of DNA Recognition and Cleavage by CRISPR-Cas9:
CRISPR-Cas9 involves base pairing between the guide RNA and the target DNA sequence, along with DNA unwinding to access both strands. The protein structure allows for precise target recognition and efficient cleavage.

Accuracy and Efficiency of CRISPR-Cas9:
CRISPR-Cas9 can identify specific DNA sequences within vast genomes. While not always 100% accurate, it demonstrates remarkable precision and efficiency.

Structural Insights into CRISPR-Cas9 Function:
3D models and structural studies reveal the intricate interactions between the protein, guide RNA, and DNA. The enzyme operates without an external energy source, relying on structural rearrangements for DNA opening and cutting.

Applications of CRISPR-Cas9:
CRISPR-Cas9 holds immense potential for addressing real-world problems across various fields. Three key areas of application include public health, agriculture, and biomedicine.

Correction of Genetic Diseases:
CRISPR-Cas9 can potentially correct disease-causing mutations, as demonstrated by the example of sickle cell anemia. This approach aims to precisely alter DNA in the tissues of patients affected by genetic disorders.

Genome Editing for Sickle Cell Disease:
Sickle cell disease is caused by a single base pair change in the DNA of a gene that produces hemoglobin. Somatic cell genome editing is a potential treatment for sickle cell disease by correcting the disease-causing mutation in blood cells. This type of editing does not affect an individual’s children, making it similar to traditional therapies.

Genome Editing in Animals for Organ Donation:
Genome editing can be used to alter pig genomes to make them better donors for organs. Two main goals of this research are to remove endogenous viral sequences and to make organs more human-like. This technology is advancing rapidly, with potential benefits for human health.

Gene Drives:
Gene drives can be used to rapidly spread a genetic trait through a population. This approach is being considered as a way to limit the spread of mosquito-borne diseases. However, there is also the potential for environmental damage, which raises ethical concerns.

Genome Editing in Agriculture:
Genome editing can be used to control the number of fruits produced by plants, alter mushroom browning, and more. This technology has sparked discussions about regulation and the definition of genetically modified organisms (GMOs). The U.S. and Europe have different stances on regulating these agricultural products.

Genome Editing for Diagnostics:
CRISPR enzymes can be used to detect DNA and RNA, enabling simple point-of-care diagnostics. This technology has the potential to detect infections and genetic variations, raising ethical questions about accessibility and responsible use.

Germline Editing in Humans:
Germline editing involves making genetic changes in eggs, sperm, or embryos, which can be passed on to future generations. This approach raises complex ethical and societal considerations, and is currently prohibited in many countries.

Importance of CRISPR-Cas9:
CRISPR-Cas9 allows humans to alter their DNA in a controlled manner, essentially altering human evolution.

CRISPR-Cas9 in Mouse Embryos:
The technology is being widely used in animal models to study diseases. Research has shown successful changes to mouse DNA that are inherited by offspring.

Ethical Concerns and the 2017 National Academies Report:
Concerns about the societal and ethical implications of CRISPR-Cas9 in the germline led to a 2017 report by the National Academies of Science. The report recommended careful analysis and public discussion before clinical use of human germline editing.

He Jiankui’s Controversial Use in Human Embryos:
He Jiankui used CRISPR-Cas9 in human embryos, resulting in the birth of twin girls with altered DNA. The stated purpose was to protect the girls from HIV infection, but the work was considered unethical and medically indefensible.

Problematic Changes in Twin Girls’ DNA:
The changes made to the DNA of the twin girls were presented as lacking scientific rigor and potentially harmful.

Technical and Ethical Concerns:
He Jiankui’s alteration of the CCR5 gene in human embryos raised concerns due to the introduction of unprecedented changes that had never been observed in the human population or tested in animals. The informed consent process and follow-up health monitoring for the children born through this experiment were poorly managed.

Potential Benefits and Future Applications:
Human germline editing holds the potential to eliminate disease-causing mutations, preventing genetic disorders like cystic fibrosis and muscular dystrophy.

International Efforts for Responsible Use:
International organizations are convening groups of scientists to address the technical, ethical, and societal questions surrounding human germline editing. The goal is to establish detailed restrictions and requirements for the clinical use of this technology.

Challenges in Regulation:
Creating a registry for all human CRISPR-related experiments is challenging due to the numerous somatic cell experiments that are ongoing. Efforts are underway to focus the registry on individuals considering genome editing in embryos for clinical applications.

Conclusion:
The rapid advancement of CRISPR-Cas9 technology emphasizes its potential as a powerful tool. Ongoing discussions aim to develop responsible guidelines and regulations for the clinical use of human germline editing.

Current and Future Challenges in Genome Editing:
Delivery remains a significant challenge in genome editing. Researchers are exploring efficient methods to deliver genome editors into cells and tissues, especially for somatic cell applications. Control over DNA changes is crucial. Scientists are working to ensure that the desired DNA alterations occur after CRISPR-Cas enzyme interactions and to minimize unintended consequences. Responsible use and regulation of genome editing technology are essential. Ethical considerations and guidelines are being developed to guide the responsible and safe application of this technology.

The Importance of Fundamental Research:
Fundamental research is the driving force behind advancements in genome editing science and technology. It lays the foundation for practical applications and enables a deeper understanding of biological processes. Collaboration between researchers, clinicians, and funding agencies is vital for advancing genome editing. Collaborative efforts foster innovation and accelerate progress.

Gratitude and Acknowledgements:
The speaker expresses gratitude to their research team, collaborators, and funding agencies, particularly the National Science Foundation, for their contributions to the field of genome editing.

Conclusion:
The speaker invites attendees to join the ongoing exploration and advancement of genome editing technology.

CRISPR: The Most Important Biological Discovery of the 21st Century:
CRISPR is not the first gene editing technology, but it is the most effective, reliable, cheap, and accurate, making it accessible to a wide range of users. Jennifer Doudna and her colleagues played a crucial role in developing CRISPR as a human tool, recognizing its immense potential and ethical implications.

CRISPR’s Ethical and Social Implications:
Jennifer Doudna has consistently expressed concerns about the ethical and social consequences of CRISPR and has actively engaged in discussions about its responsible use.

CRISPR’s Non-Human Applications:
The most significant consequences of CRISPR may lie in its non-human applications, such as improving crop yields, eradicating diseases, and conserving biodiversity. While the focus is often on human uses of CRISPR, it is important to consider the broader impact of the technology on the entire biosphere.

CRISPR and Hu Jianqui’s Fiasco:
Hank Greely disagrees with Jennifer Doudna’s characterization of Hu Jianqui as a gentleman due to his unethical and unsafe conduct in conducting human gene editing experiments.

Ethics of CRISPR in Humans:
CRISPR in humans raises ethical concerns that can be categorized along two axes: somatic vs. germline and medical vs. enhancement.

Somatic vs. Germline:
Somatic changes affect only the body cells and are not passed on to descendants, while germline changes affect both body cells and germ cells (eggs and sperm) and can be passed on to offspring.

Medical vs. Enhancement:
Medical uses of CRISPR aim to cure or prevent diseases, while enhancement uses aim to improve traits beyond the normal range, such as intelligence or athletic ability.

Four Categories of CRISPR Use:
Somatic medical: Using CRISPR to cure or prevent diseases in body cells. Somatic enhancement: Using CRISPR to enhance traits in body cells, such as improving eyesight or muscle strength. Germline medical: Using CRISPR to cure or prevent diseases in germ cells, potentially benefiting future generations. Germline enhancement: Using CRISPR to enhance traits in germ cells, potentially creating “designer babies” with desired characteristics.

Ethical Issues in Somatic Medical Use:
Safety concerns: Ensuring the safety and efficacy of CRISPR therapies. Access and equity: Ensuring that CRISPR therapies are affordable and accessible to all who need them. Prioritization of diseases: Determining which diseases to target first with CRISPR therapies.

Ethical Issues in Germline Use:
Potential for unintended consequences: Germline changes are irreversible and could have unforeseen effects on future generations. Moral and religious objections: Some people believe that germline editing is unethical or playing “God.” Concerns about eugenics: Germline editing could lead to a desire to create “designer babies” with desired traits, potentially leading to discrimination against those without those traits.

Human Germline Genome:
The human germline genome is not static but constantly changes through random mutations and human-directed changes. Human activities such as agriculture, medicine, and technology have influenced changes in the germline genome. The germline genome is not a sacred object but rather a dynamic entity that can be improved or worsened.

Germline Genome Changes:
Germline genome changes can be either beneficial or harmful. Preimplantation genetic diagnosis (PGD) is a method to select healthy embryos for implantation, avoiding diseases like cystic fibrosis. PGD is a safe and effective method that has been used to create over 15,000 healthy children.

Medical Reasons for Germline Treatment:
Germline treatment may be necessary for couples who both carry genetic diseases and want to have a healthy child. Some diseases, like Huntington’s disease, require both copies of a gene to be mutated in order to cause the disease. Couples with dominant genetic diseases cannot have healthy children from their own genes.

Ethical Concerns:
Germline fears are often overblown. Discriminating against couples who want to use germline treatment because of these fears is unfair.

Enhancement Issues:
Enhancement issues are real concerns that need to be addressed.

Ethical Axes of Consideration:
* Enhancements vs. Therapies: This is a central ethical debate, determining if CRISPR should be used to improve desirable traits or strictly for treating diseases.
* Germline vs. Somatic: Germline editing involves changes passed on to future generations, while somatic editing affects only the individual.

He Jiankui’s Experiment:
* Condemnation: Hank Greely strongly criticized He Jiankui’s germline editing experiment, calling it one of the worst he has seen.
* Need for Deterrence: Greely emphasized the importance of punishment for such actions to deter others from similar attempts.
* Transparency and Disclosure: He Jiankui’s lack of transparency and disclosure to relevant authorities was seen as a major ethical failure.

Humility and Societal Engagement:
* Scientist’s Responsibility: Greely urged scientists to be more humble and recognize that science is governed by society.
* Societal Engagement: There was a shift in the early emphasis on listening to society to a more dismissive attitude after the Hong Kong summit.

Challenges in Human In Vitro Fertilization (IVF):
* Complexity of Pre-Implantation Genetic Diagnosis (PGD): Jennifer Doudna highlighted the challenges and limitations of PGD, a technique used to select embryos free of genetic disorders.
* Rapid Advancements: Doudna emphasized the rapid progress in both IVF and genome editing technologies, requiring careful attention to their parallel development.

Balancing Risks and Benefits:
* Initial Focus on Severe Diseases: Greely suggested that germline editing should initially be used for treating severe diseases with high societal benefit.
* Long-Term Potential: In the long run, germline editing could potentially reduce the need for PGD and provide a more permanent solution for genetic disorders.

Public Policy Concerns:
* Availability of Reagents: Jennifer Doudna raised concerns about the availability of CRISPR reagents, such as Cas9 proteins, to individuals who may not have the necessary education and training to conduct experiments safely.
* Misinterpretation of Genetic Information: Doudna also highlighted the potential for individuals to misinterpret personal genetics testing results and attempt DIY gene editing without proper guidance.

Tissue Delivery:
Simply injecting a protein into the thigh is not sufficient for genome editing to work effectively. The challenge of tissue delivery arises due to the complexity of the process.

Wide Availability and Accessibility:
CRISPR genome editing is easily accessible and relatively easy to use. This raises concerns about controlling access to prevent potential misuse.

Ethical Challenges:
The widespread availability of CRISPR technology poses ethical challenges. There is a need to address potential misuse, such as unsafe or ineffective applications or unethical and dangerous uses.

Comparison to Other Technologies:
The challenge of controlling access and use is not unique to CRISPR genome editing. It applies to various technologies that are widely available, such as software engineering and hacking.

Rapid Technological Advancement:
Technologies are advancing rapidly, enabling various applications. However, regulating their use remains a challenge.

Shifting Vocabularies:
There is a shift in vocabularies between silicon-based and carbon-based life forms. Terms like “hacking” and “viruses” are used in both contexts.

Josiah Zaner as an Example:
Josiah Zaner, a Bay Area resident, injected himself with CRISPR technology. He is not considered crazy despite his unconventional actions.

DIY CRISPR Kits and Public Health:
Individuals may purchase do-it-yourself CRISPR kits for personal use, raising concerns about public health. Widespread use of such kits could lead to misuse and potential health risks if not properly regulated and understood.

Genetic Testing Misinterpretation:
Genetic testing services like 23andMe may provide incomplete or misleading results, leading to misunderstandings and misinterpretations. Individuals may receive negative test results for certain genetic conditions but still carry other dangerous versions of the gene.

Policy and Philosophical Considerations:
The ethical debate surrounding personal idiosyncratic use of CRISPR and other genetic technologies raises questions about paternalism versus free will and liberty. Accessibility and affordability of these technologies may lead to disparities in access, exacerbating healthcare inequalities.

Innovation and Societal Impact:
Scientific innovations often outpace societal understanding and regulation. Techno-innovations primarily driven by physical scientists lack input from social sciences, leading to potential societal impacts that may not be fully considered. Scientists should play a more active role in advising and implementing regulations for new technologies.

Education and Training for Ethical Use:
Training scientists to effectively communicate the significance and implications of their research to non-experts is crucial. Scientists should be encouraged to think critically about the ethical, social, legal, and political aspects of their work. Education beyond the scientific community is essential for informed decision-making and societal acceptance of new technologies.

Complexities of Genome Editing and Unintended Consequences:
Editing the genome requires extensive research and understanding of the implications to avoid unintended consequences. The genome’s current state is not sacred, but caution is needed to minimize potential negative outcomes. Both acting and not acting have consequences, and finding a balance between the two is crucial. Human culture and nature are not stable and static, so deviation from the current state is not necessarily negative. Being too cautious has its own costs, and the careful approach should consider these costs as well.

Genetic Interactions and the Human Genome:
Genetic changes in an organism can be affected by the organism’s genotype, leading to different effects in different genetic backgrounds. The same is likely true in humans, and understanding these genetic interactions is critical, especially for germline editing. Appreciation for how individual genetic changes affect humans will grow over time, initially driven by natural variants in the population. Targets for genome editing will initially be genes with evidence from natural variants, ensuring relatively confident outcomes. Predicting outcomes for genes without such information will be more challenging.

Importance of Informed Consumerism:
Being a GSA graduate or having limited biology knowledge does not prevent one from being an informed consumer of genome editing information. With effort, individuals can learn enough to make informed decisions and engage in discussions on the topic.

Common vs. Rare Alleles:
In genetic engineering, changing a common disease-causing allele to a common non-disease allele is relatively safe. Changing a common allele to a rare allele or a completely new allele raises more uncertainty and safety concerns.

CRISPR in Agriculture:
Genetically modified salmon with a single change to enhance growth and size may help solve declining fish stocks. Labeling such products as GMO may lead to unjustified rejection despite their potential benefits.

GMO Labeling:
Labeling GM products may provide consumers with desired information, but it also risks misleading or deterring them from safe and beneficial products. California’s Proposition 65 warning for carcinogens is often ignored due to its widespread use, suggesting a potential outcome for GM labeling.

International Registry vs. Local Authorities:
There is a debate on whether to create an international registry for CRISPR experiments or to trust local authorities to make ethical decisions. No definitive model has emerged yet, and the ideal approach is still under discussion.

Pre-existing Treatments vs. Genetic Solutions:
Balancing genetic interventions with existing non-genetic treatments for diseases depends on risk-benefit analysis. Genetic solutions should be weighed against well-established treatments and the severity of the condition.

HIV and Social Stigma:
In certain regions, HIV infection carries a strong social stigma. He Jiankui’s intentions to address this stigma through gene editing raised ethical questions about the motivations behind his actions.

Access to Assisted Reproduction:
China’s restrictions on assisted reproduction for HIV-positive individuals may have influenced He Jiankui’s decision to use gene editing as an alternative. The press release from the investigation suggests potential coercion and deception in recruiting participants.

Prior Knowledge and Reporting Responsibilities:
Scientists faced with knowledge of unethical research may lack clear guidelines on whom to report to or how to proceed. The need to create mechanisms for reporting and addressing questionable research practices.

Ethical Considerations and Risk-Benefit Analysis:
Ethical aspirations should include the responsibility to report unethical practices, while also providing practical mechanisms to do so. Risk-benefit analysis should be the guiding principle in balancing genetic and non-genetic treatments.

Hot, Sexy Things vs. Effective Solutions:
The allure of exciting and novel approaches can overshadow more effective but less captivating solutions. The media, the public, and the stock market often prioritize sensationalism over practical and evidence-based solutions.

Ethical Quandaries in Genetic Research:
The legality of homosexuality in China raises questions about the ethical implications of genetic engineering in different cultural contexts.

Cultural Relativity in Ethics:
Societies have varying moral codes, leading to diverse perspectives on the acceptability of genetic interventions.

Unknown Genetic Factors:
Limited knowledge about genetic determinants of traits like sexual preference and neurodiversity poses challenges in regulating genetic modifications.

Universal Human Rights:
The debate between universal human rights and cultural relativism presents a fundamental ethical dilemma.

Cultural Relativism vs. Universal Ethics:
Determining the validity of universal human rights and their origins remains a complex and unresolved issue.

Conclusion:
The intersection of genetic engineering and cultural relativism presents profound ethical challenges that require careful consideration and further exploration.