Cell culture 07
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This document appears to be the syllabus for a 7th lecture course on Advanced Animal Cell Culture offered in the 2nd semester of 2013 at Chungbuk National University's Department of Animal Science. The syllabus lists 8 topics that will be covered across 8 class dates from September to December 2013. These topics include introductions to cell culture, using cell culture as a research model, antibody/protein production in cell culture, protein production and purification, stem cells, and generating transgenic/knockout animals and genome engineering techniques. It also lists which students will present on certain topics and dates.
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Cell culture 07
- 1. 7th Lecture Advanced Aminal Cell Culture 2013 2nd Semester Department of Animal Science Chungbuk National University
- 2. Syllabus Date Topics September 5, 2013 Introduction : What is Cell Culture? September 12, 2013 Cell Culture As Model System For Research September 26, 2013 Cell Culture For Antibody / Protein Production October 17, 2013 Protein Production/Purification October 31, 2013 Stem Cell I November 14, 2013 Stem Cell II November 28, 2013 TG/KO Animals December 5, 2013 Genome Engineering/NGS December 12, 2013 Final Exam
- 3. Date September 26, 2013 Cell Culture For Antibody / Protein Production 조유미, Madhusumida October 17, 2013 Protein Production/Purification 이미진, 정용호 이영, 윤준호 October 31, 2013 November 14, 2013 Stem Cell I Jia Jia Lin, 염동현 November 28, 2013 Stem Cell II Zhao MingHui,권정우 December 5, 2013 Transgenic Animals Lin Zili, 이상배 December 12, 2013 Genome Engineering/NGS 조유진, 김상욱
- 4. Knockout/Knock In/Transgenic Animals
- 5. Why we care about that? - Knockout/Knock-In animal (mouse) : Disease Model For Medical Research - Immune-deficient Pig : Xenotransplantation? - Transgenic Animals : Animal Bioreactor? Production of useful protein in Milk or Egg? - Domestic Animal Breeding?
- 6. Knockout Mouse
- 7. Knockdown vs Knockout Gene Knockout Gene Knockdown - Expression level of mRNA was decreased by RNAi (siRNA/shRNA/dsRNA) - Gene itself are still intact - SilencingEfficiency is not ~100% - Sometime residual activities are sufficient for phenotypes - Relatively easy and fast to perform - You can do in embryonic lethal gene - Gene locus in chromosome is permanently removed/changed - Complete loss of genotype Difficulty & Time consuming Embryonic lethal gene : Conditional Knockout
- 8. Knockout mouse from Embryonic Stem Cell
- 9. Homologous Recombinantion
- 12. BAC recombineering BAC : Bacterial Artificial Chromosome Plasmid vector can have ~150kb of insert Used for the Original Human Genome Projects Ideal for modification of Large fragments for the homologus recombinantion Most of
- 13. Conditional Knockout - Why we need to make conditional knockout mouse ? Embryonic Lethal - Cre recombinase
- 15. Oocyte Specific Knockout - Cre recombinase are expressed in oocyte by ZP3 promoter - Desired locus has modified to have LoxP
- 16. Knock in Transgenic vs Knock-In?
- 17. Transgenic Animals - Embryonic Stem Cell - Somatic Nuclear Transfer - Viral Vector
- 18. Novel strategy for generation of knockout/knock-in/transgenic - Zinc-Finger Nuclease - TALEN - CRISPER Genome Engineering?
- 19. Zinc-Finger Nuclease
- 20. TALEN
- 21. CRISPR
- 22. One-step generation of Knockout Animal using TALEN