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MBB 1 August 3, 2011

From DNA to Proteins

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From DNA to Proteins

Gene unit of inheritance
A specific sequence of the DNA that is transcribed

Genetic Disorder
Dwarfism

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SCID (Severe Combined Immunodeficiency)

David Vetter (1971-1984)

Genetic Diseases
Type 1: Single locus (gene) is defective and responsible for the disease
Sickle cell anemia Hypercholesterolemia Cystic fibrosis Achondroplasia SCID (severe combined immunodeficiency)

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Genetic Diseases
Type 2: Polygenic traits may be dependent on environmental factors and lifestyle.
Heart disease Cancer Diabetes Alcoholism Schizophrenia Criminal behavior

Gene Therapy
Transfer of genetic material (DNA) for the purpose of treating diseases A normal gene is inserted into the genome to replace an abnormal, disease-causing gene

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Gene Therapy
1990 first approved gene therapy procedure W. French Anderson M.D., R. Michael Blaese, M.D., C. Bouzaid, M.D., and Kenneth Culver, M.D. Four-year old Ashanthi DeSilva born with SCID (severe combined immunodeficiency) Gene Therapy Kid

Ashanti DeSilva

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Types of Gene Therapy

Germ line gene therapy
Gene transferred into germ cells (sperm or eggs) Heritable Prohibited for application in humans

Types of Gene Therapy

Somatic cell gene therapy
Gene transferred into somatic cells (most of the body cells) Inheritable

Routes of delivery
Ex vivo cells are modified outside of the body and are transplanted back inside the body In vivo genes are changed in cells inside the body

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DNA Carriers for Gene Therapy

Can be viruses or non-viruses
Can accommodate the gene of interest Must be targeted to a limited cell type Must not be immunogenic Must be stable Can easily be produced in high concentration Can be regulated

Viral Vectors
Viruses submicroscopic particles (20-300 nm) that infect cells of a biological organism for them to reproduce
Retrovirus Adenovirus

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Viral Vectors

Viral Vectors
Naturally infectious Remove virulence genes, retain genes which allow the virus to insert its genome into its host's genome Understand the virus genetics Retrovirus or adenovirus
Retrovirus RNA virus integrates genetic material to host genome Adenovirus DNA virus does not integrate genetic material to genome

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Viral Vectors
Trouble preventing undesirable effects Problem with infecting the correct target cell in the body Making sure that the inserted gene doesn't disrupt any vital genes already in the genome

Non-Viral DNA Carriers

Advantages
Simple large-scale production Address risks using viruses

Disadvantages
Low transfection efficiency Low expression of the gene

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Non-Viral DNA Carriers

Naked DNA simplest method; DNA in plasmid or just the gene
Gene construct is made composed of the gene of interest and promoter to direct protein synthesis

Oligonucleotides synthetic
Bind to target DNA prevent expression Small RNA signal cleavage of faulty mRNA Double-stranded oligonucleotides decoys

Non-Viral DNA Carriers

Cationic liposome positively-charged lipids that can interact with negativelycharged DNA lipoplex (lipid-DNA complex) Advantages
Stable complex Can carry large-sized DNA Can target specific cells Do not induce immunological response

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Non-Viral DNA Carriers

Cationic liposome positively-charged lipids that can interact with negativelycharged DNA lipoplex (lipid-DNA complex) Disadvantages
Low transfection efficiency Shows transient expression only Shows inhibition by serum Can be toxic to certain cell types

Non-Viral DNA Carriers

Liposome Protects DNA from damage and facilitates entry into the cell

http://www.science.ca/images/Cullis_liposome.gif

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Gene Therapy

Gene Therapy Today

Severe combined immunodeficiency (SCID) Cystic fibrosis Advanced central nervous system malignancy Mesothelioma Ornithine transcarbamylase (OTC) deficiency Hemophilia Sickle cell disease

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Death from Gene Therapy

Jesse Gelsinger died in 1999 (18 years old) Suffered from OTC deficiency Used adenovirus vector infected to his liver After four days, died from massive immune response due to the virus
http://www.guineapigzero.com/images/jessephoto.jpg

Success of Gene Therapy

May 1, 2007 world's first gene therapy trial for inherited retinal disease Robert Johnson (UK) Condition: Lebers Congenital Amaurosis (LCA)

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Ethical Concerns
Pros
Cures the disease at its source / root Has great potential

Cons

Short-lived nature of gene therapy Immune response Problems with viral vectors Multigene disorders polygenic disorders Inducing a tumor Religious concerns

Other Considerations
Can this be used for cosmetic purposes
Baldness: transfer gene into follicle cells Increasing body size: growth hormone gene Increase muscle mass: dystrophin gene

In utero somatic gene therapy Should this method be used to prevent disease in addition to treating a disease?

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Perfect Humans?
Under Virginia's 1924 Eugenic Sterilization Law (based on the case of Emma, Vivian, and Carrie): 7,000 people were surgically sterilized between 1924 and 1979 because they are feebleminded, mentally defective epileptics

Gene Therapy

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