Basic Genetic Engineering-E2
|Numbering Code||U-LAS14 20036 LE68||Year/Term||2021 ・ Second semester|
|Number of Credits||2||Course Type||Lecture|
|Target Year||Mainly 1st & 2nd year students||Target Student||For science students|
|Instructor name||Adam Tsuda GUY (Graduate School of Biostudies Associate Professor)|
|Outline and Purpose of the Course||The objective of this course is to gain a familiarity with the methods, resources, and molecular tools that enable biologists to conduct their research. We will cover basic cloning strategies, expression systems, and applications that are widely used. The course is intended for 1st and 2nd year students, to provide an introduction to genetic engineering, which will serve as a foundation for more advanced courses.|
|Course Goals||Students will acquire familiarity with routine subcloning, mutagenesis, reporter constructs, epitope tags, silencing, PCR, and other important techniques. Although this is not a "wet" lab, we will learn by actually designing genetic engineering projects. Depending on enrollment, we may work in small groups or all together to plan a genetic engineering project, step by step.|
|Schedule and Contents||
Lecture topics will vary, to address the specific requirements of specific projects. I will combine short mini-lectures with in-class work in teams so that you actively learn how to use some of the design tools and strategies for genetic engineering. Lectures will vary, so as to more specifically address the actual projects that teams choose to design. Students should bring a laptop or pad with internet access so that they can work in class.
1. Overview, types of genetic engineering, biosafety. (Homework: think of ideas for genetic engineering projects. Past projects have included more nutritious rice, super-sized fugu, therapy for cachexia in cancer, DNA origami, edible vaccines, human gene therapy, arsenic decontamination, allergy-free peaches, etc. Use your imagination: the possibilities are limitless.)
2. Basic Tools: cutting and pasting. (In class, possible projects will be discussed, and students will choose about 6 of the most attractive projects, and form teams. It is possible to work alone, if so desired.)
3. Bioinformatics tools. (In class, students will work in teams to outline their project, and identify materials and steps involved.)
4. Propagating DNA constructs, plasmids and vectors. Choices for expression, editing, etc.
5. PCR, RT-PCR; primer design and amplification tips. Modern biology: DNA cloning or DNA synthesis
6. New approaches to cutting and pasting: Golden Gate, InFusion, Gateway, etc.
7. Transfection and other methods for DNA delivery into cells.
7. Making proteins visible; epitope tags, etc.
8. Tools for reverse genetics. CRISPR/Cas9 and variations
9. Knocking down genes; conditional knockouts; induction systems
10. Transgenic plants, mice, fish…
11. Lecture topic tailored to specific projects
12. Lecture topic tailored to specific projects
13. Lecture topic tailored to specific projects
14. Final Exam (first set of group presentations)
15. Final Exam (second set of group presentations)
16. Feedback Class
|Evaluation Methods and Policy||Final grades will be based on quizzes (10%), a final presentation (30%), and attendance and participation (60%).|
|Course Requirements||The course is designed for 1st and 2nd year students, from all science backgrounds. Recombineering is conceptually not difficult. The vocabulary will get a bit technical, but with some effort, non-biology students should understand most of the course. Students should bring a laptop or pad with internet access so that they can work in class.|
|Study outside of Class (preparation and review)||
As we get into individual or team cloning projects, some outside reading or planning may be necessary, roughly 1-2 hours per week.
Students may also need to spend some time working on their independent projects outside of class, although about half of each class will be spent on the group projects.
|Textbooks||Textbooks/References||I will teach you how to use many online sources and freeware to work with DNA sequences, vectors, cloning, etc.|
|References, etc.||Current Protocols in Molecular Biology (Kyoto University library online subscription). URL: http://onlinelibrary.wiley.com/book/10.1002/0471142727/toc|