CRISPR: From Nature to Bench and Bedside
Continuing Education Credits
Objectives
- Describe the discovery of CRISPR and its role as an elegant bacterial immune defense system.
- State the fundamental components of the CRISPR system from nature.
- Explain the basics of a lab-modified CRISPR/Cas9 gene correction tool for hereditary medical conditions (i.e., sickle cell).
- Explain how CRISPR can improve the xenograft organ transplant.
- Discuss CRISPR medical ethics and the corresponding challenges and future perspectives.
Course Outline
- CRISPR Discovery
- CRISPR Discovery
- What is CRISPR?
- CRISPR is a unique type of immune system first discovered in:
- Bacteria-invading viruses are called:
- CRISPR is a discreetly organized system that serves as the storage bank of representative sequence information. A CRISPR system is composed of all of the following except:
- CRISPR From Nature: Structure and Function
- Structure and Function
- How Does the CRISPR System Work?
- How Does the CRISPR System Work, continued
- True or False: Phage DNA is obtained out of bacterial phage by the host bacterium and stored in the CRISPR locus as interspacers.
- Regarding interspacers, select the accurate statement from the choices provided.
- During the initial phage invasion of bacteria, phage DNA is shuffled to the bacterial CRISPR locus. Which unique phage DNA does CRISPR select?
- CRISPR Characterization and Engineering
- CRISPR Characterization and Engineering
- CRISPR Characterization and Engineering: Dr. Doudna and Dr. Charpentier
- CRISPR Characterization and Engineering: Dr. Zhang
- CRISPR Characterization and Engineering: Dr. Church and Dr. Liu
- True or False: Cas9 was purified from mammalian cells.
- How is Cas9 cutting action regulated?
- The successful purification of Cas9 was first performed by Dr. Doudna and Dr. Charpentier. The two bacteria they used to isolate purified Cas9 were:
- CRISPR/Cas9 Associated DNA Repair Pathways
- CRISPR/Cas9 Associated DNA Repair Pathways
- CRISPR/Cas9 Associated DNA Repair Pathways: Two Major Types of Repairs
- True or False: Non-homologous end joining (NHEJ) is the preferred cellular DNA repair mechanism that fixes DNA breakage (blunt ends on both DNA strands).
- Knock-in of a new DNA segment is carried out by:
- Which of the following statements is correct about homology-directed repair (HDR)?
- Select the accurate statement regarding CRISPR/Cas9 gene silencing.
- CRISPR Gene Editing and Sickle Cell Disease
- CRISPR/Cas9 in the Clinic: Sickle Cell Disease
- Sickle Cell Disease
- Discovery of BCL11A Gene
- Casgevy Preparation Procedures
- Manufacturing Price Tag
- True or False: CRISPR/Cas9-mediated silencing of the BCL11A gene also silenced the fetal hemoglobin gene.
- Casgevy preparation procedures include all of the following, except:
- CRISPR and Xenograft Organ Transplantation
- CRISPR/Cas9 in the Clinic: Xenograft Organ Transplantation
- Host Immune Rejection
- Host Immune Rejection, continued
- True or False: Genes that encode sugar groups on the antigens expressed on the surface of pig cells are edited out by CRISPR/Cas9 aimed at reducing host immune rejection.
- All of the following were utilized in the first pig heart transplant, except:
- True or False: The removal of the selected genes is a work in progress in terms of efficacy assessment. More research is required to gauge suitability and effectiveness.
- Ethical Issues
- Ethical Issues and CRISPR Gene Editing - A True Story
- Overview of HIV Infection
- HIV Targeting and Making Genetic Alterations in Embryos
- True or False: Adult stem cells taken from a patient and subjected to CRISPR/Cas9 gene alteration affect only the patient undergoing treatment but will not be inherited and passed on to future generations.
- All of the following statements regarding HIV infection are true, except:
- Dr. He Jiankui used the CRISPR/Cas9 gene editing tool to edit wild-type CCR5 in two human embryos, subsequently giving rise to two baby girls. What was the subsequent findings?
- CRISPR Challenges, Reflections, and Future Perspectives
- CRISPR Challenges and Limitations
- Reflections: Approaches
- Reflections: Approaches, continued
- Reflections: DNA Sequencing
- Further Reflection and Future Perspectives
- True or False: Overcoming CRISPR's off-target effect is key to ensuring CRISPR's staying power as a gene editing tool.
- The best approach to ensuring that the intended DNA has been successfully excised, leading to silencing or knockdown of the intended target, is to do what?
- All of the following were discussed as promising tools to alleviate CRISPR challenges, except:
- References
- References
Additional Information
Level of Instruction: Intermediate to advanced
Intended Audience: Medical laboratory scientists, medical
laboratory technicians, microbiologists, and molecular biologists. This
course is also appropriate for MLS and MLT students, molecular biology students, and pathology residents.
Author Information: Dr. Nancy Liu-Sullivan served as a Senior Research Scientist with a specialty in cancer genomics and drug discovery before joining the biology faculty at the College of Staten Island (CSI), City University of New York (CUNY), teaching Immunology, Radiation Biology, and General Biology, in addition to mentoring students in cancer research. Dr. Liu-Sullivan is also the author of MediaLab’s CE courses titled "Hallmarks and Signaling of Cancer Cell" and "HLA and Cancer Immunotherapy".
Reviewer Information:
Dr. Julie Ann West is certified by the American Society for Clinical Pathology (ASCP) as a Medical Laboratory Scientist (MLS) and as a Specialist in Microbiology (SM). In addition, Dr. West has earned a PhD in Public Health, Epidemiology Specialization (emphasis on infectious disease), and is Certified in Public Health (CPH) by the National Board of Public Health Examiners. Dr. West is experienced as a Technical Specialist, Safety Officer, Educator, and Lead in the Veterans Administration Healthcare System and has prior experience as an Administrative Laboratory Director.
Ralph Garippa, Ph.D., is the Director of Gene Editing and RNAi Core Facility at Memorial Sloan Kettering Cancer Center, with more than two decades of drug discovery expertise and leadership. He is the former Head of Cell-Based High Throughput Screening and Microscopic Imaging-based High Content Screening at Hoffmann-La Roche’s Nutley, NJ facility. His previous academic partners included Harvard University, Massachusetts General Hospital, and The Hebrew University in Jerusalem, Israel. Dr. Garippa holds a Ph.D. in Pharmacology from Columbia University and a B.A. degree in Biology from Fairleigh Dickinson University.
Course Description: This course describes the discovery, structure, and function of CRISPR DNA sequences, DNA repair pathways, and gene editing. It also discusses associated ethical issues, challenges, and future perspectives.