Introduction of Biotechnology
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1 生物技术导论 Introduction of Biotechnology 肖卫华教授 Tel: (O); 30053(L) Web: 助教 : 姚立超 : ylc@mail.ustc.edu.cn Spring 2018 谢思琪 :xiesq@mail.ustc.edu.cn
2 生物技术导论 - 课程安排 Introduction of Biotechnology
3 成绩考核标准 课堂讲座 ( 考核 )50% 100% 课堂纪律 ( 不定时点名 ) 30% 缺席一次总成绩扣 10 分 提问 (2 次 ) 20% 每次提问加 10 分 截止日期 : 第一次 :3 月 24 日 24:00 第二次 :4 月 28 日 24:00
4 课堂讲座 : 总分 100 分, 占最终总成绩 40% 题目 : 请根据目前对人类重大疾病基本理论的认识和了解, 如传染病 肿瘤 心血管疾病 老年性疾病等, 制定一个相关的研究计划 研究计划内容包括 : 1. 研究背景和存在问题 ( 阅读相关的研究文献, 发现和提出待解决的问题 ) 2. 研究目的 ( 如病原体致病性机制 ; 新诊断产品和方法 ; 新预防措施 ; 治疗靶点或治疗方法等 研究 ) 3. 总体研究方案 ( 围绕科学问题和研究目的, 采用体外 体内 动物模型 临床标本或患者等 开展系统性和逻辑性的研究规划 ) 4. 技术路线和选用的实验方法 ( 具体实施研究方案, 包括具体的实验材料和方法 ) 5. 预期结果 ( 可能得到的研究结论, 回答研究题目提出的科学问题 ). 研究的创新点 ( 研究思路 手段, 研究结果的意义等 ); 讲座要求 : 1. 每位 ( 组 ) 同学独立准备一份 PPT 演讲稿并在课堂上讲述, 中英文均可, 不能互相抄袭 ; 2. 讲座时间为 5-8 分钟 ( 根据选课人数待定 ), 每超时 1 分钟扣 5 分 ; 3. 评分标准将参考以下几个方面 :(1) 完整 ( 包括上述 1- 项 );(2) 科学性和逻辑性 ( 依据充分 );(3) 演讲能力 ( 语言 趣味 );(4) ppt 制作质量 ( 美观 图文并茂 )
5 Transgenic 5 Core Technologies Genetic Engineering
6 5 Core Technologies Genetic Engineering What is Genetic Engineering? Genetic engineering refers to a set of technologies that are being used to change the genetic makeup of cells and move genes across species boundaries to produce novel organisms. The techniques involve highly sophisticated manipulations of genetic material and other biologically important chemicals. Normal Mice Knockout Mice Natural Corn Transgene Corn
7 5 Core Technologies Genetic Engineering The overview of Genetic Engineering
8 5 Core Technologies Genetic Engineering Gene Targeting (Knockout) Transgene (Knock-in) Normal Mice Knockout Mice Natural Corn Transgene Corn
9 5 Core Technologies Genetic Engineering Workflow of Gene Targeting Recombinant and Selection
10 5 Core Technologies Genetic Engineering Techniques Genetic Engineering Recombinant DNA, Introducing rdna into ES and Selection electro- and chemical transportation, microinjection, bioballistics Reimplantation Breed Selection
11 5 Core Technologies Genetic Engineering Transgene (Knock-in) Gene Targeting (Knockout)
12 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering Homology Recombination
13 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering Homology Recombination
14 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering Nuclease-mediated Targeting Nat. review Genetics
15 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering Science 339, 78 (2013); Triplex-forming Oligonucleotides
16 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering site-specific nucleases vs. Homology recombinant Overview of possible genome editing outcomes using site-specific nucleases.
17 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering Nuclease-mediated Targeting- ZFNs
18 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering Nuclease-mediated Targeting- TALENs Nat. review Genetics
19 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering CRISPR-Cas9
20 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering CRISPR-Cas9
21 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering CRISPR-Cas9
22 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering Diagram of the possible mechanism for CRISPR
23 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering TALEN
24 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering TALEN
25 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering Model for DNA target specificity of TAL effectors.
26 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering Repeats of TAL effectors and predicted target sequences
27 Trends in Biotechnology July 2013, Vol. 31, No. 7 Structure of zinc-finger and transcription activator-like effectors (TALEs).
28 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering ZFN vs. TALEN
29 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering
30 5 Core Technologies Genetic Engineering Basic Principle of genetic engineering Nat. review Genetics
31 5 Core Technologies Genetic Engineering Techniques Genetic Engineering Recombinant DNA, Introducing rdna into ES and Selection electro- and chemical poration, microinjection, bioballistics Reimplantation Breed Selection
32 5 Core Technologies Genetic Engineering Transgenic In animal
33 5 Core Technologies Genetic Engineering Recombination and Selection
34 5 Core Technologies Genetic Engineering Recombination and Selection Recombination and Selection Cytotxic to cells
35 5 Core Technologies Genetic Engineering Recombination and Selection
36 5 Core Technologies Genetic Engineering Transgene Animal Red Mouse Fluorescent Protein
37 5 Core Technologies Genetic Engineering Transgenic in Plant - Overview
38 5 Core Technologies Genetic Engineering Transgenic - In Plant Pesticides Herbicides
39 5 Core Technologies Genetic Engineering Transgenic - In Plant
40 5 Core Technologies Genetic Engineering Transgenic - In Plant Transgene Plant
41 5 Core Technologies Genetic Engineering The Risk of Transgene Escape for Agriculture Pros Transgenic crops are often environmentally friendly; Farmers can use fewer pesticides. Cons Effect on non-target organisms, such as insects, in the food web; Movement of herbicide resistance to weeds.
42 5 Core Technologies Genetic Engineering Transgene Animal Advantage Salmon Non-transgenic salmon AquAdvantage Salmon Siblings at ca months after first-feeding
43 5 Core Technologies Genetic Engineering Transgene Animal Muscle Calf Double-muscle phenomenon is result of natural mutation in Myostatin coding sequence November 1997
44 5 Core Technologies Genetic Engineering Transgene Animal Cassie Line Enviropig TM Three months Five months
45 5 Core Technologies Genetic Engineering Transgene Animal Human Antibody Production Calves carrying human antibody genes. Calves produce specific human antibody after immunization Microchromosome with human antibody genes in cow cell
46 5 Core Technologies Genetic Engineering Transgene Animal Green Chicken Lentiviral vectors can be used to generate transgenic lines with an efficiency in the order of 100-fold higher than any previously published method Source: Efficient production of germline transgenic chickens using lentiviral vectors, McGrew et al., 2004 EMBO reports VOL 5, NO 7
47 5 Core Technologies Genetic Engineering Promises of Genetic Engineering produce new medicines and safer vaccines. treat some genetic diseases. enhance biocontrol agents in agriculture. increase agricultural yields and decrease production costs. decrease allergy-producing characteristics of some foods. improve food nutritional value. develop biodegradable plastics. decrease water and air pollution. slow food spoilage. control viral diseases. inhibit inflammation.
48 Core Technologies Stem Cell & Cloning
49 Core Technologies Stem Cell & Cloning What is Stem Cell? Stem cell A cell that has the ability to divide (self replicate) for indefinite periods in culture and to give rise to specialized cells. Two Properties: Ability to Differentiate Ability to Self-regenerate Three Processes: Development Repair of damaged tissue Cancer resulting
50 Core Technologies Stem Cell & Cloning Types of Stem Cells According to the source Embryo Stem Cells Adult stem cell (Somatic Stem Cells) Stage of Differentiations
51 Core Technologies Stem Cell & Cloning Sources of Stem Cells Embryo Adult Stem Cells from Various Tissues
52 Core Technologies Stem Cell & Cloning Stem Cell in Mammalian Development 全能干细胞 多能干细胞 多能干细胞 定向干细胞 定向干细胞 分化细胞 Hierarchy of stem cells during differentiation at each stage, differential potential decreases and specialization increases.
53 Core Technologies Stem Cell & Cloning Stem Cell In Mammalian Development Early in development: Later in development: Type-I Later in development: Type II
54 Core Technologies Stem Cell & Cloning Plasticity of adult stem cell (Somatic Stem Cell) Adult stem cell An undifferentiated cell found in a differentiated tissue that can renew itself and (with certain limitations) differentiate to yield all the specialized cell types of the tissue from which it originated. Plasticity Property: Increase in differentiation potential of a cell Roles: Replacement of damaged and injured tissue
55 Core Technologies Stem Cell & Cloning Adult stem cell in Tissue Repair Tissue regeneration is achieved by two mechanisms: (1) Circulating stem cells divide and differentiate under appropriate signaling by cytokines and growth factors, e.g. blood cells; and (2) Differentiated cells which are capable of division can also self-repair, e.g. hepatocytes, endothelial cells, smooth muscle cells, keratinocytes and fibroblasts.
56 Core Technologies Stem Cell & Cloning Cancer Stem Cells Cancer stem cells have been identified in blood, brain, and breast cancers, and more recently in ovarian, colon and pancreas cancers. The new findings provide further support for the stem cell hypothesis, the theory that some tumors contain small populations of self-renewing cells that give rise to all of the cells in the tumor.
57 Core Technologies Stem Cell & Cloning Stem Cell Technology Stem Cell Cultivation
58 Core Technologies Stem Cell & Cloning Stem Cell Technology Nuclear Transfer Nuclear Transfer Techniques 1. Enucleate Embryo Cell Somatic cell Nuclear Donor 2. Microinject 3. Culture Cloned embryo 2. or Electrical Pulse
59 Core Technologies Stem Cell & Cloning Stem Cell Technology Nuclear Transfer Microinjection Enucleate Microinject
60 Core Technologies Stem Cell & Cloning Fertilization vs. Nuclear Transfer
61 Core Technologies Stem Cell & Cloning ips Induced Pluripotent Stem Cell
62 Core Technologies Stem Cell & Cloning Applications of Stem Cell Technology Not allowed! Developing
63 Core Technologies Stem Cell & Cloning Category of Cloning Reproductive cloning:creating a new organism from a single cell of an adult. The genes (but not the far less-important mitochondrial DNA) of the offspring are identical to the parent. Therapeutic cloning: creating embryos to supply embryonic stem cells as a source of spare parts to treat disease.
64 Core Technologies Stem Cell & Cloning Applications for Reproductive Cloning in Animal Dolly was born on 5 July 199
65 Core Technologies Stem Cell & Cloning Applications for Reproductive Cloning in Animal Arthritis (AR) -- a progressive degeneration of bones and joints -- at a mere five years of age. And yet that's what's happened to Dolly, the first mammal made by nuclear transfer, commonly called cloning.
66 Core Technologies Stem Cell & Cloning Applications for Reproductive Cloning in Animal Animal cloning:jersey Calf Clones University of Tennessee
67 Core Technologies Stem Cell & Cloning Applications for therapeutic Cloning in Animal Pluripotent, embryonic stem cells originate as inner mass cells with in a blastocyst. The stem cells can become any tissue in the body, excluding a placenta. Only the morula's cells are totipotent, able to become all tissues and a placenta.
68 Core Technologies Stem Cell & Cloning Embryo Stem Cell for Therapeutic Cloning in Human
69 Core Technologies Stem Cell & Cloning ips For Therapeutic
70 Core Technologies Stem Cell & Cloning Development of Stem Cells
71 Developmental Regulation of Hematopoiesis in the Mouse
72 Core Technologies Stem Cell & Cloning Hematopoietic Development in the Zebrafish
73 Core Technologies Stem Cell & Cloning Development of Stem Cells
74 Core Technologies Stem Cell & Cloning Development of Stem Cells With increasing age the clonal composition within the HSC compartment changes. A proportional shift in lineage potential from balanced to myeloidbiased is observed. Moreover, while there is an increase in HSC number, the per-cell quality of HSCs decreases with age.
75 Core Technologies Stem Cell & Cloning Development of Stem Cells Aging of the hematopoietic system
76 Core Technologies Stem Cell & Cloning Development of Stem Cells
77 Core Technologies Stem Cell & Cloning Development of Stem Cells Overview of proposed age-associated micro-environmental (extrinsic) and intrinsic changes of the hematopoietic system: first, increase in the number of phenotypically defined HSCs; second, more distant localization of HSCs from the endosteum; third, less supportive stroma; fourth, accumulation of adipocytes in the bone marrow; fifth, different cytokine milieu; sixth, increased DNA-damage; seventh, increased exposure to reactive oxygen species (ROS); and eighth, changes in gene expression and accumulation of epigenetic alterations.
78 Core Technologies Stem Cell & Cloning Development of Stem Cells
79 Core Technologies Stem Cell & Cloning Development of Stem Cells. The haematopoietic tree
80 Core Technologies Stem Cell & Cloning Development of Stem Cells Haematopoietic stem cell niches
81 Core Technologies Stem Cell & Cloning Development of Stem Cells Haematopoietic stem cell niches
82 Core Technologies Stem Cell & Cloning Regulation of Stem Cell Development Possible mechanisms by which bone marrow niches adapt to changes
83 Core Technologies Stem Cell & Cloning Dynamic behaviors of HSC in response to injury
84 Core Technologies Stem Cell & Cloning homeostatic HSCs
85 Core Technologies Stem Cell & Cloning In response to loss of HSC
86 Core Technologies Stem Cell & Cloning During the repair of HSC
87 Core Technologies Stem Cell & Cloning During the recovery of HSC,
88 Core Technologies Stem Cell & Cloning Dynamic behaviors of HSC in response to injury
89 Core Technologies Stem Cell & Cloning Dynamic behaviors of HSC in response to injury
90 Core Technologies Stem Cell & Cloning Interferon promotes cell cycle entry of dormant and homeostatic haematopoietic stem cells
91 Leukemia stem cells may be targeted by a two-step strategy.
92 Core Technologies Stem Cell & Cloning. The push and pull on HSCs. The push : haematopoietic stem cells (HSCs) divide in direct response to stimuli. The pull : HSCs divide following the depletion of committed progenitor populations from the bone marrow
93 Core Technologies Stem Cell & Cloning Immunomodulation of HSCs during infection
94 Core Technologies Stem Cell & Cloning. Inflammation as a rheostat for haematopoietic stem cell differentiation
95 Core Technologies Stem Cell & Cloning Immune cell activation, cell states and network
96 Core Technologies Stem Cell & Cloning
97
98 Core Technologies Stem Cell & Cloning Transcription Factor Antagonism in Lineage Determination
99 Core Technologies Stem Cell & Cloning Reprogramming of Hematopoietic Lineages
100 Core Technologies Stem Cell & Cloning Epigenetics Regulations in HSC development Heritable epigenetic mechanisms
101 Core Technologies Stem Cell & Cloning Epigenetics Regulations in HSC development Epigenetic switches in myeloid versus lymphoid cells.
102 Core Technologies Stem Cell & Cloning Epigenetics Regulations in B Cell Develoment Choreography of allelic choice in B cells
103 Core Technologies Stem Cell & Cloning mirnas in HSC Develoment mirnas in hematopoiesis and immune system
104 mirnas in HSC Development Key ES cell mirnas associated with ES pluripotency, selfrenewal, differentiation.
105 Core Technologies Stem Cell & Cloning mirnas in HSC Develoment Key mirnas at the center of ips and CSC
106 Core Technologies ips Human Skin cell Oct3/4, Sox2, c-myc, Klf4 or Oct4 only in a neural cell ips
107 Contributors of Stem Cell and ips Albert Lasker Basic Medical Research Award The Albert Lasker Award for Basic Medical Research is one of the prizes awarded for the understanding, diagnosis, prevention, treatment, and cure of disease. Almost 50% of the winners have gone on to win Nobel prize. The announcement will be hold on Oct 2, 2009 Sir John Bertrand Gurdon Wolf Prize in Medicine (1989) Albert Lasker Basic Medical Research Award (2009) Nobel Priz (2012) Shinya Yamanaka Robert Koch Prize (2008) Shaw Prize (2008) Albert Lasker Basic Medical Research Award (2009) Nobel Priz (2012)
108 Tumorgenesis from Stem Cell in Chronic Inflammation
109 Inflammation-associated Carcinogenesis Step1-DNA damage (NO) (TNF- -> NF- B) Step2-Proliferation (TGF-b, EGF, IGFs)
110 Core Technologies Stem Cell & Cloning Cloning Ethic challenges Cloning Isn't as Easy as It Sounds...
111
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