Authors Rahul K. Keswani, Mihael Lazebnik & Daniel W. Pack
|
|
- Winifred Owen
- 6 years ago
- Views:
Transcription
1 Reinstatement of Retroviral Infectivity via Non-Covalent Attachment of DOTAP, DOPE and Cholesterol to Murine Leukemia Virus-Like Particles Hybrid Viral/Lipid Gene Delivery Vectors Authors Rahul K. Keswani, Mihael Lazebnik & Daniel W. Pack Laboratory of Advanced Drug and Gene Delivery Department of Chemical and Biomolecular Engineering University of Illinois, Urbana-Champaign AIChE Annual Meeting 2011 October 18, 2011 Session: Nucleic Acid Delivery II 1
2 Overview Introduction & Background Why hybrid vectors? What are RVLPs? Results Transfections Uptake Sizes Morphology Conclusions and Future Work 2
3 Gene Therapy ASGCT defines Gene Therapy as The treatment of disease by either replacing damaged or abnormal genes with normal ones, or by providing new genetic instructions to help fight disease. Retrovirus Adenovirus Adeno-associated Virus Plasmid Polymer Liposomes Polymers Polyethylenimine (PEI) Poly-L-Lysine (PLL) Polyamidoamine (PAMAM) Chitosan Lipids Cationic transfection lipids (DOTAP;DC-Chol) Neutral Helper lipids (DOPE;Cholesterol) Sources:
4 Comparison Why do we need a hybrid design? Retrovirus Synthetic Vector Transfection Efficiency Excellent Poor-excellent Size Small Small-medium Gene Expression Stable Transient Toxicity No Yes/No Flexible Targeting No Yes Pathogenicity Yes Unknown/No Immunogenicity Yes Little Production Expensive Inexpensive Stability Poor Robust 4
5 Retrovirus Structure Protease Lipid bilayer Envelope Protein Reverse Transcriptase Integrase Icosahedral capsid shell RNA (2 copies) 5
6 Retrovirus Envelope Protein needs a suitable alternative Envelope protein provides good efficiency but Immunogenicity Pathogenicity Poor Stability Poor Processing Single Tropism Retrovirus-Like Particle (RVLP) 6
7 Brief Protocol Synthetic envelopes are added non-covalently to RVLPs GP293Luc Transfect with plasmid encoding for envelope protein Vortex and Incubate for 2-4 h Quantify via Real-Time PCR assay 7
8 Synthetic Envelopes This study improves the feasibility of the hybrid vector design Generation 1 LF2000 PEI PLL Generation 2 Chitosan DOTAP/DOPE/Cholesterol Scale bar = 100 nm Ramsey et al, 2010, Journal of Controlled Release 8
9 Synthetic Envelopes Lipids (φ) are efficient non-viral gene delivery vectors Cationic Lipid 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) Helper Lipids (Neutral) 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) Cholesterol 9
10 φ-rvlps Transfection Efficiency is dependent on the lipid composition 10
11 φ-rvlps Transfection Efficiency is dependent on the lipid composition 11
12 φ-rvlps Transfection Efficiency is dependent on the lipid composition 12
13 φ-rvlps Transfection Efficiency is dependent on the lipid composition 13
14 φ-rvlps Transfection Efficiency is dependent on the lipid composition (DOTAP-heavy) and (DOTAP-light,DOPE-heavy) lipid compositions performed worse DOPE and Cholesterol, if present in approximately equal proportions, was optimal On repeat trials, DOTAP:DOPE:Cholesterol = 5:8:7 (w) best composition for consistently good transfection efficiency 14
15 Measuring Cellular Uptake in HEK293 cells RVLPs labeled with DiD prior to formation of hybrid vectors + Lipophilic fluorescent dye 15
16 φ-rvlps Cationic charge on lipids improves uptake but has no effect on transfection 5 μg lipid/10 9 RVLPs 16
17 φ-rvlps Cationic charge on lipids improves uptake but has no effect on transfection 10 μg lipid/10 9 RVLPs 17
18 φ-rvlps Cationic charge on lipids improves uptake but has no effect on transfection 20 μg lipid/10 9 RVLPs 18
19 Normalized Metabolic Activity Lipid Toxicity 505 was toxic 10 µg/ml to HEK293 cells Lipo550 Lipo587 Lipo Lipids (µg/ml) 19
20 φ-rvlps TEM of hybrid vectors shows enveloping of RVLP 20
21 Size Reduction for φ-rvlps Extrusion of lipids led to smaller sizes of hybrid vectors Simple Hydration Type Only Lipid 5* 10* Lipo ± ± ± 60 Lipo ± ± ± 7 Lipo ± ± ± 184 Sizes in nm; * - µg/10 9 RVLPs After Extrusion Type Only Lipid 5* 10* Lipo ± ± ± 10 Lipo ± ± ± 5 Lipo ± ± ± 8 Sizes in nm; * - µg/10 9 RVLPs 21
22 Size Reduction for φ-rvlps TEM scans of hybrid vectors formed by extrusion method 587 alone 22
23 Size Reduction for φ-rvlps TEM scans of hybrid vectors formed by extrusion method Lipo µg/10 9 RVLPs 23
24 Normalized RLU/mg total protein Size Reduction for φ-rvlps Normalized RLU/mg total protein Normalized RLU/mg total protein Extruded lipid hybrid vectors had mixed performance in transfections 4.5 Lipo Lipo505 Lipo550e 3.5 Lipo505e Lipid (μg)/10 9 RVLPs Lipid (μg)/10 9 RVLPs Lipo Lipo587e Lipid (μg)/10 9 RVLPs 24
25 Conclusions and Future Work Optimized lipid composition for lipid-based hybrid vectors in HEK293 cells Uptake has no correlation with subsequent transfection within the cells Size reduction of hybrid vector aggregates successfully achieved via lipid extrusion Formation of smaller liposomes led to smaller hybrid vectors Synthetic envelope decides the intracellular navigation route? - Amphotropic Virus = Lipo-RVLPs > Polymer-RVLPs (10-fold difference) Future Work Compare the intracellular trafficking of lipid-based hybrid vectors with chitosan-based hybrid vectors and amphotropic virus Evaluate receptor-targeted gene delivery and serum performance via PEGylated hybrid vectors 25
26 Acknowledgements Professor Daniel W. Pack (Advisor) Colleagues: Joshua, Neel, Steve, Nate, Dave, Lily, Ti-An, Shy Chyi, Kalena, Kara, Victor, Mark, Yujie, Mihael, Noel Undergrads Mikhil, Andreina, Janelle, Ian, Hao Facilities Institute of Genomic Biology (Real-Time PCR, Confocal Microscopy) Beckman Institute (Confocal Microscopy) Roy J. Carver Biotechnology Center (Real-Time PCR, Flow Cytometry) Center for Microscopic Imaging Lou Ann Miller (Transmission Electron Microscopy) Frederick Seitz Materials Research Laboratory (Transmission Electron Microscopy) Cell Media Facility (Sandy McMasters) Funding National Science Foundation National Institutes of Health Hanratty Travel Award Graduate College Travel Award Biotechnology Center, UIUC (Catherine Connor Outstanding Dissertation Award) 26
27 Comparison Why do we need a hybrid design? Retrovirus Hybrid Vector Synthetic Vector Transfection Efficiency Excellent Poor-Excellent Poor-excellent Size Small Medium-Large Small-medium Gene Expression Stable Stable 1 Transient Toxicity No Yes/No Yes/No Flexible Targeting No Yes 2 Yes Pathogenicity Yes Unknown Unknown/No Immunogenicity Yes Little Little Production Expensive Cheaper Inexpensive Stability Poor Robust Robust 1 Depends on synthetic envelope 2 Untested 27
28 28
29 Real-Time PCR Assay Protocol RNA Isolation Reverse Transcription Real-Time PCR 29
30 Real-Time PCR Assay Quantification 30
31 31
Oncolytic Adenovirus Complexes Coated with Lipids and Calcium Phosphate for Cancer Gene Therapy
Oncolytic Adenovirus Complexes Coated with Lipids and Calcium Phosphate for Cancer Gene Therapy Jianhua Chen, Pei Gao, Sujing Yuan, Rongxin Li, Aimin Ni, Liang Chu, Li Ding, Ying Sun, Xin-Yuan Liu, Yourong
More informationRecombinant Protein Expression Retroviral system
Recombinant Protein Expression Retroviral system Viruses Contains genome DNA or RNA Genome encased in a protein coat or capsid. Some viruses have membrane covering protein coat enveloped virus Ø Essential
More informationGene Therapies. Josh Crockett, Lexi Trujillo, Caitlin Halliday
Gene Therapies Josh Crockett, Lexi Trujillo, Caitlin Halliday Gene Therapy Main objective: create carriers that can envelope and deliver genetic materials into target tissues (like cancerous cells) in
More informationChapter 19: Viruses. 1. Viral Structure & Reproduction. What exactly is a Virus? 11/7/ Viral Structure & Reproduction. 2.
Chapter 19: Viruses 1. Viral Structure & Reproduction 2. Bacteriophages 3. Animal Viruses 4. Viroids & Prions 1. Viral Structure & Reproduction Chapter Reading pp. 393-396 What exactly is a Virus? Viruses
More informationChapter 19: Viruses. 1. Viral Structure & Reproduction. 2. Bacteriophages. 3. Animal Viruses. 4. Viroids & Prions
Chapter 19: Viruses 1. Viral Structure & Reproduction 2. Bacteriophages 3. Animal Viruses 4. Viroids & Prions 1. Viral Structure & Reproduction Chapter Reading pp. 393-396 What exactly is a Virus? Viruses
More informationSECTION 25-1 REVIEW STRUCTURE. 1. The diameter of viruses ranges from about a. 1 to 2 nm. b. 20 to 250 nm. c. 1 to 2 µm. d. 20 to 250 µm.
SECTION 25-1 REVIEW STRUCTURE VOCABULARY REVIEW Define the following terms. 1. virus 2. capsid 3. retrovirus 4. viroid 5. prion MULTIPLE CHOICE Write the correct letter in the blank. 1. The diameter of
More informationViruses. Picture from:
Viruses Understand the structure of bacteriophages & human immunodeficiency virus (HIV) Appreciate that viruses replicate in host cells (thereby destroying them) Picture from: http://eands.caltech.edu/articles/lxvii1/viruses.html
More informationWeek 5 Section. Junaid Malek, M.D.
Week 5 Section Junaid Malek, M.D. HIV: Anatomy Membrane (partiallystolen from host cell) 2 Glycoproteins (proteins modified by added sugar) 2 copies of RNA Capsid HIV Genome Encodes: Structural Proteins
More informationACQUIRED IMMUNODEFICIENCY SYNDROME AND ITS OCULAR COMPLICATIONS
ACQUIRED IMMUNODEFICIENCY SYNDROME AND ITS OCULAR COMPLICATIONS Acquired immunodeficiency syndrome (AIDS ) is an infectious disease caused by a retrovirus, the human immunodeficiency virus(hiv). AIDS is
More informationVIROLOGY. Engineering Viral Genomes: Retrovirus Vectors
VIROLOGY Engineering Viral Genomes: Retrovirus Vectors Viral vectors Retrovirus replicative cycle Most mammalian retroviruses use trna PRO, trna Lys3, trna Lys1,2 The partially unfolded trna is annealed
More informationViral reproductive cycle
Lecture 29: Viruses Lecture outline 11/11/05 Types of viruses Bacteriophage Lytic and lysogenic life cycles viruses viruses Influenza Prions Mad cow disease 0.5 µm Figure 18.4 Viral structure of capsid
More informationFayth K. Yoshimura, Ph.D. September 7, of 7 RETROVIRUSES. 2. HTLV-II causes hairy T-cell leukemia
1 of 7 I. Diseases Caused by Retroviruses RETROVIRUSES A. Human retroviruses that cause cancers 1. HTLV-I causes adult T-cell leukemia and tropical spastic paraparesis 2. HTLV-II causes hairy T-cell leukemia
More informationStructure & Function of Viruses
Structure & Function of Viruses Discovery of Viruses Louis Pasteur- looks for a causative agent for rabies, says too small, can not find it 1892 Dimitry Ivanosky- studies tobacco disease, can not find
More informationHepatitis B Antiviral Drug Development Multi-Marker Screening Assay
Hepatitis B Antiviral Drug Development Multi-Marker Screening Assay Background ImQuest BioSciences has developed and qualified a single-plate method to expedite the screening of antiviral agents against
More informationJulianne Edwards. Retroviruses. Spring 2010
Retroviruses Spring 2010 A retrovirus can simply be referred to as an infectious particle which replicates backwards even though there are many different types of retroviruses. More specifically, a retrovirus
More informationOverview: Chapter 19 Viruses: A Borrowed Life
Overview: Chapter 19 Viruses: A Borrowed Life Viruses called bacteriophages can infect and set in motion a genetic takeover of bacteria, such as Escherichia coli Viruses lead a kind of borrowed life between
More informationWHY? Viruses are considered non-living because they do:
Viruses What is a Virus? Non-living particle WHY? Viruses are considered non-living because they do: NOT Carry out metabolism NOT Grow or develop NOT Replicate without the help of a living cell (host).
More information11/15/2011. Outline. Structural Features and Characteristics. The Good the Bad and the Ugly. Viral Genomes. Structural Features and Characteristics
Chapter 19 - Viruses Outline I. Viruses A. Structure of viruses B. Common Characteristics of Viruses C. Viral replication D. HIV II. Prions The Good the Bad and the Ugly Viruses fit into the bad category
More information19 Viruses BIOLOGY. Outline. Structural Features and Characteristics. The Good the Bad and the Ugly. Structural Features and Characteristics
9 Viruses CAMPBELL BIOLOGY TENTH EDITION Reece Urry Cain Wasserman Minorsky Jackson Outline I. Viruses A. Structure of viruses B. Common Characteristics of Viruses C. Viral replication D. HIV Lecture Presentation
More informationRecent Advancements in Virus Detection and Mechanistic Fate. Krista Wigginton Assistant Professor of Environmental Engineering University of Michigan
Recent Advancements in Virus Detection and Mechanistic Fate Krista Wigginton Assistant Professor of Environmental Engineering University of Michigan Acknowledgments for the graduate students who conducted
More informationVirology Introduction. Definitions. Introduction. Structure of virus. Virus transmission. Classification of virus. DNA Virus. RNA Virus. Treatment.
DEVH Virology Introduction Definitions. Introduction. Structure of virus. Virus transmission. Classification of virus. DNA Virus. RNA Virus. Treatment. Definitions Virology: The science which study the
More informationAntiviral Drugs Lecture 5
Antiviral Drugs Lecture 5 Antimicrobial Chemotherapy (MLAB 366) 1 Dr. Mohamed A. El-Sakhawy 2 Introduction Viruses are microscopic organisms that can infect all living cells. They are parasitic and multiply
More informationThis training module is required for all personnel listed on an IBC protocol that describes work utilizing viral vectors (both replication competent
This training module is required for all personnel listed on an IBC protocol that describes work utilizing viral vectors (both replication competent and incompetent) regardless of the biosafety level used
More informationTransfection mechanisms of polyplexes, lipoplexes. DLD-1 colorectal cancer cells
Supporting Information Transfection mechanisms of polyplexes, lipoplexes and stealth liposomes in α 5 β 1 integrin bearing DLD-1 colorectal cancer cells Maroof M. Adil, Zachary S. Erdman and Efrosini Kokkoli*
More informationSupplementary Information. Supplementary Figure 1
Supplementary Information Supplementary Figure 1 1 Supplementary Figure 1. Functional assay of the hcas9-2a-mcherry construct (a) Gene correction of a mutant EGFP reporter cell line mediated by hcas9 or
More informationVIRUSES. 1. Describe the structure of a virus by completing the following chart.
AP BIOLOGY MOLECULAR GENETICS ACTIVITY #3 NAME DATE HOUR VIRUSES 1. Describe the structure of a virus by completing the following chart. Viral Part Description of Part 2. Some viruses have an envelope
More informationLight and X-ray triggered liposomal gene/drug delivery system for cancer therapy
Light and X-ray triggered liposomal gene/drug delivery system for cancer therapy Wei Deng, 1 Wenjie Chen, 2 Sandhya Clement, 1 Anna Guller, 1 Alexander Engel, 3 Ewa Goldys 1 1 The Graduate School of Biomedical
More informationSupplemental information contains 7 movies and 4 supplemental Figures
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 Supplemental information contains 7 movies and 4 supplemental Figures Movies: Movie 1. Single virus tracking of A4-mCherry-WR MV
More information19/06/2013. Viruses are not organisms (do not belong to any kingdom). Viruses are not made of cells, have no cytoplasm, and no membranes.
VIRUSES Many diseases of plants and animals are caused by bacteria or viruses that invade the body. Bacteria and viruses are NOT similar kinds of micro-organisms. Bacteria are classified as living organisms,
More information19 2 Viruses Slide 1 of 34
1 of 34 What Is a Virus? What Is a Virus? Viruses are particles of nucleic acid, protein, and in some cases, lipids. Viruses can reproduce only by infecting living cells. 2 of 34 What Is a Virus? Viruses
More informationReoviruses. Virion. Genome. Genes and proteins. Viruses and hosts. Diseases. Distinctive characteristics
Reoviruses Virion Genome Genes and proteins Viruses and hosts Diseases Distinctive characteristics Virion Naked icosahedral capsid (T=13), diameter 60-85 nm Capsid consists of two or three concentric protein
More informationRabies virus-like particles expressed in HEK293 cells
Engineering Conferences International ECI Digital Archives Vaccine Technology IV Proceedings Spring 5-21-2012 Rabies virus-like particles expressed in HEK293 cells Diego Fontana Cell Culture Laboratory
More informationDATA SHEET. Provided: 500 µl of 5.6 mm Tris HCl, 4.4 mm Tris base, 0.05% sodium azide 0.1 mm EDTA, 5 mg/liter calf thymus DNA.
Viral Load DNA >> Standard PCR standard 0 Copies Catalog Number: 1122 Lot Number: 150298 Release Category: A Provided: 500 µl of 5.6 mm Tris HCl, 4.4 mm Tris base, 0.05% sodium azide 0.1 mm EDTA, 5 mg/liter
More informationHybrid Lipid-Coated Silver Nanoparticles for Drug Delivery. Sven Burke and Marilyn R. Mackiewicz, Ph.D.
Hybrid Lipid-Coated Silver Nanoparticles for Drug Delivery. Sven Burke and Marilyn R. Mackiewicz, Ph.D. 1 What is the driving force behind our research? 2 Men Women Cancer is 2 nd deadliest disease in
More informationOctober 26, Lecture Readings. Vesicular Trafficking, Secretory Pathway, HIV Assembly and Exit from Cell
October 26, 2006 Vesicular Trafficking, Secretory Pathway, HIV Assembly and Exit from Cell 1. Secretory pathway a. Formation of coated vesicles b. SNAREs and vesicle targeting 2. Membrane fusion a. SNAREs
More informationVirology. *Viruses can be only observed by electron microscope never by light microscope. The size of the virus: nm in diameter.
Virology We are going to start with general introduction about viruses, they are everywhere around us; in food; within the environment; in direct contact to etc.. They may cause viral infection by itself
More informationChapter 6- An Introduction to Viruses*
Chapter 6- An Introduction to Viruses* *Lecture notes are to be used as a study guide only and do not represent the comprehensive information you will need to know for the exams. 6.1 Overview of Viruses
More informationNucleic acid: singled stranded, double stranded, RNA, or DNA, linear or circular. Capsid: protein coat that is most of the mass of the virus.
Viruses General Characteristics of Viruses 1. Depending on view may be regarded as exceptionally complex aggregates of nonliving chemicals or as exceptionally simple living microbes. 2. Contain a single
More informationLipid Bilayer Based Binding Surfaces for Nucleic Acids
Lipid Bilayer Based Binding Surfaces for Nucleic Acids Caitlin Kreutz Major: Biology Mentor: Emin Oroudjev Faculty Advisor: Helen Hansma Funding By: National Institutes of Health July 28 th, 2005 Main
More informationIdentification of Microbes Lecture: 12
Diagnostic Microbiology Identification of Microbes Lecture: 12 Electron Microscopy 106 virus particles per ml required for visualization, 50,000-60,000 magnification normally used. Viruses may be detected
More informationChoosing Between Lentivirus and Adeno-associated Virus For DNA Delivery
Choosing Between Lentivirus and Adeno-associated Virus For DNA Delivery Presenter: April 12, 2017 Ed Davis, Ph.D. Senior Application Scientist GeneCopoeia, Inc. Outline Introduction to GeneCopoeia Lentiviral
More informationRetroviruses. ---The name retrovirus comes from the enzyme, reverse transcriptase.
Retroviruses ---The name retrovirus comes from the enzyme, reverse transcriptase. ---Reverse transcriptase (RT) converts the RNA genome present in the virus particle into DNA. ---RT discovered in 1970.
More informationViral Genetics. BIT 220 Chapter 16
Viral Genetics BIT 220 Chapter 16 Details of the Virus Classified According to a. DNA or RNA b. Enveloped or Non-Enveloped c. Single-stranded or double-stranded Viruses contain only a few genes Reverse
More informationLESSON 4.6 WORKBOOK. Designing an antiviral drug The challenge of HIV
LESSON 4.6 WORKBOOK Designing an antiviral drug The challenge of HIV In the last two lessons we discussed the how the viral life cycle causes host cell damage. But is there anything we can do to prevent
More information5/6/17. Diseases. Disease. Pathogens. Domain Bacteria Characteristics. Bacteria Viruses (including HIV) Pathogens are disease-causing organisms
5/6/17 Disease Diseases I. II. Bacteria Viruses (including HIV) Biol 105 Chapter 13a Pathogens Pathogens are disease-causing organisms Domain Bacteria Characteristics 1. Domain Bacteria are prokaryotic.
More information8/13/2009. Diseases. Disease. Pathogens. Domain Bacteria Characteristics. Bacteria Shapes. Domain Bacteria Characteristics
Disease Diseases I. Bacteria II. Viruses including Biol 105 Lecture 17 Chapter 13a are disease-causing organisms Domain Bacteria Characteristics 1. Domain Bacteria are prokaryotic 2. Lack a membrane-bound
More informationChapter 18. Viral Genetics. AP Biology
Chapter 18. Viral Genetics 2003-2004 1 A sense of size Comparing eukaryote bacterium virus 2 What is a virus? Is it alive? DNA or RNA enclosed in a protein coat Viruses are not cells Extremely tiny electron
More informationVirus Basics. General Characteristics of Viruses. Chapter 13 & 14. Non-living entities. Can infect organisms of every domain
Virus Basics Chapter 13 & 14 General Characteristics of Viruses Non-living entities Not considered organisms Can infect organisms of every domain All life-forms Commonly referred to by organism they infect
More informationSupporting Information for. Uptake of poly(2-hydroxypropylmethacrylamide)-coated gold nanoparticles
Supporting Information for Uptake of poly(2-hydroxypropylmethacrylamide)-coated gold nanoparticles in microvascular endothelial cells and transport across the blood-brain barrier Christian Freese #*, Ronald
More informationViral structure م.م رنا مشعل
Viral structure م.م رنا مشعل Viruses must reproduce (replicate) within cells, because they cannot generate energy or synthesize proteins. Because they can reproduce only within cells, viruses are obligate
More informationVIROLOGY PRINCIPLES AND APPLICATIONS WILEY. John B. Carter and Venetia A. Saunders
VIROLOGY PRINCIPLES AND APPLICATIONS John B. Carter and Venetia A. Saunders WILEY -urief Contents Preface to Second Edition xix Preface to First Edition xxi Abbreviations Used in This Book xxiii Greek
More informationVirus Basics. General Characteristics of Viruses 5/9/2011. General Characteristics of Viruses. Chapter 13 & 14. Non-living entities
Virus Basics Chapter 13 & 14 General Characteristics of Viruses Non-living entities Not considered organisms Can infect organisms of every domain All life-formsf Commonly referred to by organism they infect
More informationViral Vectors In The Research Laboratory: Just How Safe Are They? Dawn P. Wooley, Ph.D., SM(NRM), RBP, CBSP
Viral Vectors In The Research Laboratory: Just How Safe Are They? Dawn P. Wooley, Ph.D., SM(NRM), RBP, CBSP 1 Learning Objectives Recognize hazards associated with viral vectors in research and animal
More informationChapter 13 Viruses, Viroids, and Prions. Biology 1009 Microbiology Johnson-Summer 2003
Chapter 13 Viruses, Viroids, and Prions Biology 1009 Microbiology Johnson-Summer 2003 Viruses Virology-study of viruses Characteristics: acellular obligate intracellular parasites no ribosomes or means
More informationAP Biology. Viral diseases Polio. Chapter 18. Smallpox. Influenza: 1918 epidemic. Emerging viruses. A sense of size
Hepatitis Viral diseases Polio Chapter 18. Measles Viral Genetics Influenza: 1918 epidemic 30-40 million deaths world-wide Chicken pox Smallpox Eradicated in 1976 vaccinations ceased in 1980 at risk population?
More informationLESSON 4.4 WORKBOOK. How viruses make us sick: Viral Replication
DEFINITIONS OF TERMS Eukaryotic: Non-bacterial cell type (bacteria are prokaryotes).. LESSON 4.4 WORKBOOK How viruses make us sick: Viral Replication This lesson extends the principles we learned in Unit
More informationInfluenza virus exploits tunneling nanotubes for cell-to-cell spread
Supplementary Information Influenza virus exploits tunneling nanotubes for cell-to-cell spread Amrita Kumar 1, Jin Hyang Kim 1, Priya Ranjan 1, Maureen G. Metcalfe 2, Weiping Cao 1, Margarita Mishina 1,
More informationMicrobiology. Microbiology
Microbiology Microbiology What are GERMS? What are GERMS? Microorganisms that make you sick (pathogens) There are many different types of microorganisms: Bacteria (strep throat, food poisoning like E.
More informationPhytanyl substituted asymmetric gemini surfactant-based transfection vectors for gene therapy
Phytanyl substituted asymmetric gemini surfactant-based transfection vectors for gene therapy by Haitang Wang A thesis presented to the University of Waterloo in fulfillment of the thesis requirement for
More informationLEC 2, Medical biology, Theory, prepared by Dr. AYAT ALI
General Characteristics, Structure and Taxonomy of Viruses Viruses A virus is non-cellular organisms made up of genetic material and protein that can invade living cells. They are considered both a living
More informationA novel N-acyl phosphatidylethanolamine-containing delivery vehicle for spermine-condensed plasmid DNA
NONVIRAL TRANSFER TECHNOLOGY (2000) 7, 769 783 2000 Macmillan Publishers Ltd All rights reserved 0969-7128/00 $15.00 www.nature.com/gt RESEARCH ARTICLE A novel N-acyl phosphatidylethanolamine-containing
More informationRama Nada. - Malik
- 2 - Rama Nada - - Malik 1 P a g e We talked about HAV in the previous lecture, now we ll continue the remaining types.. Hepatitis E It s similar to virus that infect swine, so its most likely infect
More informationModified Liposomes with Biomaterials Delivery Applications
Modified Liposomes with Biomaterials Delivery Applications Dr. Rahau Shirazi ACS Meeting Newcastle, Australia November 8th, 2013 utline Motivation and Background Focus Future Direction Liposomes Delivery
More informationSupplementary Material
Supplementary Material Nuclear import of purified HIV-1 Integrase. Integrase remains associated to the RTC throughout the infection process until provirus integration occurs and is therefore one likely
More informationI. Bacteria II. Viruses including HIV. Domain Bacteria Characteristics. 5. Cell wall present in many species. 6. Reproduction by binary fission
Disease Diseases I. Bacteria II. Viruses including are disease-causing organisms Biol 105 Lecture 17 Chapter 13a Domain Bacteria Characteristics 1. Domain Bacteria are prokaryotic 2. Lack a membrane-bound
More informationENDOCYTOSIS MECHANISM OF POLY- AND LIPOPLEXES IN SIRNA DELIVERY IN HELA CELLS MIHAEL LAZEBNIK THESIS
ENDOCYTOSIS MECHANISM OF POLY- AND LIPOPLEXES IN SIRNA DELIVERY IN HELA CELLS BY MIHAEL LAZEBNIK THESIS Submitted in partial fulfillment of the requirements for the degree of Master of Science in Chemical
More informationModule 2 In vivo gene therapy. Lecture 7. In-situ, in-vivo and ex-vivo gene therapy (part I)
Module 2 In vivo gene therapy Lecture 7 In-situ, in-vivo and ex-vivo gene therapy (part I) Somatic cell gene therapy involves the transfer of gene to a diseased somatic cell either within the body or outside
More informationSupplementary Fig. 1. Delivery of mirnas via Red Fluorescent Protein.
prfp-vector RFP Exon1 Intron RFP Exon2 prfp-mir-124 mir-93/124 RFP Exon1 Intron RFP Exon2 Untransfected prfp-vector prfp-mir-93 prfp-mir-124 Supplementary Fig. 1. Delivery of mirnas via Red Fluorescent
More informationLahore University of Management Sciences. BIO314 Virology and Microbiology (Spring 2015)
BIO314 Virology and Microbiology (Spring 2015) Instructor Room. Office Hours Email Telephone Secretary/TA TA Office Hours Course URL (if any) Shaper Mirza and Sadia Hamera Shaper.Mirza@uth.tmc.edu Course
More informationLecture Readings. Vesicular Trafficking, Secretory Pathway, HIV Assembly and Exit from Cell
October 26, 2006 1 Vesicular Trafficking, Secretory Pathway, HIV Assembly and Exit from Cell 1. Secretory pathway a. Formation of coated vesicles b. SNAREs and vesicle targeting 2. Membrane fusion a. SNAREs
More informationCurrent Strategies in HIV-1 Vaccine Development Using Replication-Defective Adenovirus as a Case Study
Note: I have added some clarifying comments to the slides -- please click on Comments under View to see them. Current Strategies in HIV-1 Vaccine Development Using Replication-Defective Adenovirus as a
More informationDoctoral Degree Program in Marine Biotechnology, College of Marine Sciences, Doctoral Degree Program in Marine Biotechnology, Academia Sinica, Taipei,
Cyclooxygenase 2 facilitates dengue virus replication and serves as a potential target for developing antiviral agents Chun-Kuang Lin 1,2, Chin-Kai Tseng 3,4, Yu-Hsuan Wu 3,4, Chih-Chuang Liaw 1,5, Chun-
More informationA ph-dependent Charge Reversal Peptide for Cancer Targeting
Supporting Information A ph-dependent Charge Reversal Peptide for Cancer Targeting Naoko Wakabayashi 1, Yoshiaki Yano 1, Kenichi Kawano 1, and Katsumi Matsuzaki 1 1 Graduate School of Pharmaceutical Sciences,
More informationSome living things are made of ONE cell, and are called. Other organisms are composed of many cells, and are called. (SEE PAGE 6)
Section: 1.1 Question of the Day: Name: Review of Old Information: N/A New Information: We tend to only think of animals as living. However, there is a great diversity of organisms that we consider living
More informationLESSON 1.4 WORKBOOK. Viral sizes and structures. Workbook Lesson 1.4
Eukaryotes organisms that contain a membrane bound nucleus and organelles. Prokaryotes organisms that lack a nucleus or other membrane-bound organelles. Viruses small, non-cellular (lacking a cell), infectious
More informationOptimization of the Fuse-It-mRNA Protocol for L929 Cells in the µ-plate 24 Well
Optimization of the Fuse-It-mRNA Protocol for L929 Cells in the µ-plate 24 Well 1. General Information... 1 2. Background... 1 3. Material and Equipment Required... 2 4. Experimental Procedure and Results...
More informationHIV & AIDS: Overview
HIV & AIDS: Overview UNIVERSITY OF PAPUA NEW GUINEA SCHOOL OF MEDICINE AND HEALTH SCIENCES DIVISION OF BASIC MEDICAL SCIENCES DISCIPLINE OF BIOCHEMISTRY & MOLECULAR BIOLOGY PBL SEMINAR VJ TEMPLE 1 What
More informationL I F E S C I E N C E S
1a L I F E S C I E N C E S 5 -UUA AUA UUC GAA AGC UGC AUC GAA AAC UGU GAA UCA-3 5 -TTA ATA TTC GAA AGC TGC ATC GAA AAC TGT GAA TCA-3 3 -AAT TAT AAG CTT TCG ACG TAG CTT TTG ACA CTT AGT-5 OCTOBER 31, 2006
More informationVirion Genome Genes and proteins Viruses and hosts Diseases Distinctive characteristics
Hepadnaviruses Virion Genome Genes and proteins Viruses and hosts Diseases Distinctive characteristics Hepatitis viruses A group of unrelated pathogens termed hepatitis viruses cause the vast majority
More informationThe chemical fate of biological pollutants in treatment processes
The chemical fate of biological pollutants in treatment processes http://pubs.rsc.org/en/content/articlehtml/2011/sm/c1sm060 92a http://www.epa.gov/nerlcwww/polio.htm Source: Centers for Disease Control
More informationFig. 1: Schematic diagram of basic structure of HIV
UNIVERSITY OF PAPUA NEW GUINEA SCHOOL OF MEDICINE AND HEALTH SCIENCES DIVISION OF BASIC MEDICAL SCIENCES DISCIPLINE OF BIOCHEMISTRY & MOLECULAR BIOLOGY PBL SEMINAR HIV & AIDS: An Overview What is HIV?
More information1. Virus 2. Capsid 3. Envelope
VIRUSES BIOLOGY II VOCABULARY- VIRUSES (22 Words) 1. Virus 2. Capsid 3. Envelope 4. Provirus 5. Retrovirus 6. Reverse transcriptase 7. Bacteriophage 8. Lytic Cycle 9. Virulent 10. Lysis 11. Lysogenic Cycle
More informationSUPPORTING INFORMATION
SUPPORTING INFORMATION SUPPLEMENTARY FIGURE LEGENDS Fig. S1. Separation of non-dissolved nanoparticles. Tests were conducted on the separation of non-dissolved nanoparticles added in excess to BEGM (A)
More information7.012 Quiz 3 Answers
MIT Biology Department 7.012: Introductory Biology - Fall 2004 Instructors: Professor Eric Lander, Professor Robert A. Weinberg, Dr. Claudette Gardel Friday 11/12/04 7.012 Quiz 3 Answers A > 85 B 72-84
More informationB19, see Parvovirus B19 Bone marrow, gene transfer with parvovirus. Erythrovirus, see Parvovirus B19, Simian parvovirus
... Subject Index Adeno-associated virus Cap and genome encapsidation 87 DNA integration homologous recombination 90, 91 latency vs replication 77, 78 mechanism 79 requirements 78, 79 site in human genome
More informationHIV INFECTION: An Overview
HIV INFECTION: An Overview UNIVERSITY OF PAPUA NEW GUINEA SCHOOL OF MEDICINE AND HEALTH SCIENCES DIVISION OF BASIC MEDICAL SCIENCES DISCIPLINE OF BIOCHEMISTRY & MOLECULAR BIOLOGY PBL MBBS II SEMINAR VJ
More information3. on T helper {cells / lymphocytes} ; 3. ACCEPT macrophages / dendritic cells / CD4 cells
1(a) 1. (structure G is {glycoprotein / gp120} ; 2. used for {attachment / eq} to CD4 (molecules / receptors /antigens) ; 1. IGNORE gp 41 and gp 160 and other wrong numbers 3. on T helper {cells / lymphocytes}
More informationGeneral Virology I. Dr Esam Ibraheem Azhar (BSc, MSc, Ph.D Molecular Medical Virology) Asst. Prof. Medical Laboratory Technology Department
General Virology I Dr Esam Ibraheem Azhar (BSc, MSc, Ph.D Molecular Medical Virology) Asst. Prof. Medical Laboratory Technology Department ١ General Virology I Lecture Outline Introduction istory Definition
More informationSize nm m m
1 Viral size and organization Size 20-250nm 0.000000002m-0.000000025m Virion structure Capsid Core Acellular obligate intracellular parasites Lack organelles, metabolic activities, and reproduction Replicated
More informationChapter 13B: Animal Viruses
Chapter 13B: Animal Viruses 1. Overview of Animal Viruses 2. DNA Viruses 3. RNA Viruses 4. Prions 1. Overview of Animal Viruses Life Cycle of Animal Viruses The basic life cycle stages of animal viruses
More informationToward the Safe Use of Lentivirus and Retrovirus Vector Systems
27 July, 2017 The Asian Conference on Safety & Education in Laboratory Toward the Safe Use of Lentivirus and Retrovirus Vector Systems Takaomi Sanda, MD, PhD Principal Investigator, Cancer Science Institute
More informationQuantifying Lipid Contents in Enveloped Virus Particles with Plasmonic Nanoparticles
Quantifying Lipid Contents in Enveloped Virus Particles with Plasmonic Nanoparticles Amin Feizpour Reinhard Lab Department of Chemistry and the Photonics Center, Boston University, Boston, MA May 2014
More informationDr. Ahmed K. Ali Attachment and entry of viruses into cells
Lec. 6 Dr. Ahmed K. Ali Attachment and entry of viruses into cells The aim of a virus is to replicate itself, and in order to achieve this aim it needs to enter a host cell, make copies of itself and
More informationSupplemental Materials and Methods Plasmids and viruses Quantitative Reverse Transcription PCR Generation of molecular standard for quantitative PCR
Supplemental Materials and Methods Plasmids and viruses To generate pseudotyped viruses, the previously described recombinant plasmids pnl4-3-δnef-gfp or pnl4-3-δ6-drgfp and a vector expressing HIV-1 X4
More informationMicroRNA sponges: competitive inhibitors of small RNAs in mammalian cells
MicroRNA sponges: competitive inhibitors of small RNAs in mammalian cells Margaret S Ebert, Joel R Neilson & Phillip A Sharp Supplementary figures and text: Supplementary Figure 1. Effect of sponges on
More informationSupplementary Figure 1. SC35M polymerase activity in the presence of Bat or SC35M NP encoded from the phw2000 rescue plasmid.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 Supplementary Figure 1. SC35M polymerase activity in the presence of Bat or SC35M NP encoded from the phw2000 rescue plasmid. HEK293T
More informationBIT 120. Copy of Cancer/HIV Lecture
BIT 120 Copy of Cancer/HIV Lecture Cancer DEFINITION Any abnormal growth of cells that has malignant potential i.e.. Leukemia Uncontrolled mitosis in WBC Genetic disease caused by an accumulation of mutations
More informationRevisiting the Definition of Living Thing
Biology of Viruses (Ch 0 p77 and 88-9) What do you already know about viruses? Revisiting the Definition of Living Thing How did we define a living thing? H0 DOMAIN ARCHAEA virus So, if the Cell Theory
More informationViruses Tomasz Kordula, Ph.D.
Viruses Tomasz Kordula, Ph.D. Resources: Alberts et al., Molecular Biology of the Cell, pp. 295, 1330, 1431 1433; Lehninger CD Movie A0002201. Learning Objectives: 1. Understand parasitic life cycle of
More information