Basics of CRISPR/Cas System

Students will be able to:

• Understand the CRISPR/Cas system as a natural adaptive immune system in prokaryotes.

• Explain the mechanism of CRISPR/Cas, including how the guide RNA (gRNA) directs Cas9 to specific genetic sequences to induce double-strand breaks.

• Identify the role of cellular repair mechanisms such as Non-Homologous End Joining (NHEJ) and Homology Directed Repair (HDR) in gene editing.

• Discuss the basic principles of genome editing and how CRISPR is used to modify or delete specific genes.

Students will be able to:

• Apply knowledge of CRISPR as a tool for genome editing, specifically its precision and site-specific targeting.

• Understand CRISPR applications in biomedical research, such as gene knockout, gene silencing, and gene knock-in techniques.

• Analyze CRISPR’s role in agricultural biotechnology, including how it can be used for crop improvement and disease resistance.

• Explore CRISPR applications in biotechnology, such as creating genetically modified organisms (GMOs) and advancing drug development and gene therapies.

Students will be able to:

• Trace the historical development of CRISPR, from its initial discovery in 1987 to its current role as a genome-editing tool.

• Recognize key milestones in the evolution of CRISPR, including its identification in prokaryotes, understanding its function in bacterial defense, and its use in gene editing.

• Understand the major advancements in CRISPR technology, such as the discovery of Cas12a, RNA editing, and its application in human embryos.

• Appreciate the ethical considerations and major achievements, such as the Nobel Prize in Chemistry awarded to Doudna and Charpentier for their CRISPR discoveries.

Students will have gained a foundational understanding of the CRISPR/Cas system and will be able to:

• Understand the basic structure of CRISPR technology

Students will have gained a foundational understanding of the CRISPR/Cas system and will be able to:

• Identify and describe the key components of the CRISPR/Cas system, including tracrRNA, crRNA, gRNA and Cas proteins.

• Understand the function of the Ribonucleoprotein Complex (RNP) in gene editing.

  
  

Students will have gained a foundational understanding of the CRISPR/Cas system and will be able to:

• Identify and describe the key components of the CRISPR/Cas system, including protospacers and PAM sequences.

• Understand the function of the Target genes in gene editing.

• Discuss the importance of protospacers and PAM sequences in ensuring precise DNA targeting during CRISPR/Cas9 activity.

Students will be able to:

• Describe different Classes & Types of CRISPR/Cas system.

Students will be able to:

• Describe different Classes and Type 1 & 3 of CRISPR/Cas system.

Students will be able to:

• Describe different Classes and Type 2 of CRISPR/Cas system.

• Describe CRISPR/Cas9 system.

Students will be able to:

• Describe different Classes and Type 2 of CRISPR/Cas system.

• Describe CRISPR/Cas 4,5 and 6 system.

Students will be able to:

• Identify the different types and variants of Cas proteins and understand their specific roles in CRISPR technology.

• Explain the primary differences between Cas proteins that target DNA versus those that target RNA.

• Differentiate between natural and engineered Cas proteins and describe their significance in reducing off-target effects.

• Analyze the specific characteristics of Cas proteins.

By the end of this lecture, students will be able to:

• Describe the three Phases of the CRISPR/Cas mechanism: Adaptation, crRNA Expression, and Target Interference.

• Explain how the Adaptation phase stores viral DNA as a memory to enhance bacterial defense.

• Understand the role of crRNA in guiding the Cas protein for precise targeting and cleavage of invading viral DNA.

By the end of this lecture, students will be able to:

• Explain the 5 steps of adaptation, including fragmentation, trimming, and integration of DNA into the CRISPR locus.

• Understand the significance of the Protospacer Adjacent Motif (PAM) in the selection and integration of protospacers.

• Describe the role of Cas1 and Cas2 in facilitating nucleophilic attack and ligation during spacer integration.

By the end of this lecture, students will be able to:

• Understand the process of crRNA biogenesis, including the conversion of pre-crRNA to mature crRNA through the action of Cas proteins or RNase III.

• Explain the role of mature crRNAs in guiding Cas proteins to target foreign DNA during interference.

• Describe the mechanism of Double-Strand Break (DSB) during target interference, including the role of PAM sequences and Cas proteins in cleaving foreign DNA.

• Understand the different delivery methods of CRISPR/Cas, including in vitro and in vivo techniques, and their applications in gene editing.

• Evaluate the strengths and limitations of each delivery method in terms of efficiency, precision, and applicability to research or therapeutic contexts.

• Differentiate between viral and non-viral delivery systems for CRISPR/Cas components and their respective advantages and challenges.

• Assess the factors influencing the choice of delivery method, such as safety, efficiency, immune response, and target tissue specificity.

• Understand the two main DNA repair pathways after CRISPR-induced breaks: Non-Homologous End Joining (NHEJ) and Homology Directed Repair (HDR).

• Determine which repair mechanism to use based on the goal of the experiment, whether it's gene knockout via NHEJ or precise gene correction using HDR.

By the end of this lecture, students will be able to:

• Understand how CRISPR/Cas technology is applied to enhance desirable traits in fish, such as muscle growth and disease resistance, through specific gene modifications.

• Explain the role of CRISPR in controlling invasive species by manipulating fertility or environmental sensitivity to reduce their impact on ecosystems.

By the end of this lecture, students will be able to:

• Identify the key limitations of CRISPR/Cas technology, including ethical concerns, off-target effects, and high costs.

• Discuss the challenges specific to CRISPR applications in aquaculture, such as the small size of fish eggs and the lack of genomic resources for aquatic species.