______ ______ is The manipulation of an individuals genomic information to produce a therapeutic outcome.
Gene Therapy
First human gene therapy trial started in 1990 with the correction of an ______ ______.
inherited disease (SCID)
Of the clinical trials sfrom 1990-1999, how many were curative? Definitive?
None
Jesse Gelsinger had _____ _____ deficiency
ornithine transcarbamylase
Jess Gelsinger died as a result of an immune response to a ______ _____
adenociral vector
Jolee Mohr died as a result of an immune response to a _______
Adeno Associated Virus
In which of the following ways is NOT a way one can intervene to treat acquired diseases?
A) Gene augmentation therapy
B) Eliminate selected cell populations
C) Correct an acquired gene mutation
D) Gene transfection with cationic liposomes
All are possible interventions?
this approach involves the introduction of a single gene to overcome loss of gene function.
Gene Augmentation Therapy
A requirement of gene augmentation therapy is?
High expression of the replacement gene
The Elimintation of Selected Cells generally involves the insertion of a specific gene or code to produce what?
A cellular toxin
Specificity is important in the Elimination of Selected Cell approach. Why?
all cells inserted with the gene of interesst will be killed
to protect cells from inadvertant gene toxicity one can insert a _____ _____ instead fo a toxin gene.
Prodrug gene
in Prodrug activation therapy, gangciclovir is monophosphorylated by _____ _____ in tumor cells infected with HSV genes .
in the Bystander Effect, adjacent cells become transfected with _____ _____
ganciclovir monophosphate
gap junctions between tumor cells are created by _____ ______
connexon proteins
The E. coli cytosine deaminase
5-fluorocytosine ? 5 fluorouracil system has been used to do what?
substitution of gene for one that has been altered due to injury is known as what thype of gene therapy?
Correction of a Gene Mutation
Correction of a Gene Mutation requires _____ _____ to replace damaged genes with wild-types
homologous recombination
A zinc finger nuclease is an engineered fusion protein containing what 2 propeties?
1) DNA cleavage domain of the restriction endonuclease Fokl 2) Zf DNA binding domain
Fokl needs to _____ for activity, so use two nonpalindromic proteins
dimerize
homologous recombination occurs in the A) zinc finger domain B) nuclease domain
nuclease
the HIV - CCR5 gene knockout used a A) ex vivo B) in vivo approach.
ex vivo
Ad Van Tages to Ex Vivo therapy include: A) many cell populations are exposed to therapy B) cells numbers can be expanded to large amounts C) only selecting cells with transferred gene reduces success.
B. Only one cell pop is exposed (advantage) and only selecting cells that have been transfected is an advantage
Dis Ad Van Tages to Ex Vivo therapy include A)Must have access to the cells of interest B) Tx may require more cells than can be produced C) lack of target cell specificity
A & B (C is a disadvantage of in vivo)
In Vivo advantages include all BUT:
D) (C can be a benefit or a disadvantage)
The primary disadvantage with In Vivo therapy is?
Targeting the gene of interest to a specific cell type.
Th need to get stable integration of a gene into the target cell if one wants to achieve long term effects is the _____ _____ of gene therapy.
Essential Requirement
Techniques to introduce DNA into Human cells includes all BUT: A) viral vectors B) Goldmember's Gold Plated, Gold shooting, Gene Gun C) Receptor-mediated exocytosis D) Cationic polymer complexes
C. ENDOcytosis. (OK maybe it isn't Goldmember's, but gene guns loaded with gold are pretty standard issue)
Why can't you use retroviral vectors in vivo?
because complement kills retrovirus
Retroviral Vectors infect _____ cells.
dividing
Dhing Question: This is an effective system that has a high efficiency of gene transfer and viral integration. A) Retro viral vector B) Gene Gun C) Cationic Polymers D) liposomes
A (the others may be true, but this is from the slide)
This virus has a natural affinity for airway epithelium, cornea and gut (receptor dependent).
Adenoviral Vector
Adenoviral vectors infects ____ Cells
non-dividing
Adenoviral Vector uses ______ mediated endocytosis for cell uptake
receptor
Adenoviral Vector virus grows to high/low titers for good infectivity
high
T/F Adenoviral Vectors Do not stably integrate into the host cell genomic DNA, so only good for transient transfection of the cells.
TRUE
Adenoviral Vectors Produce an inflammatory response by the body which precludes this system from many applications.
TRUE
Advantages of Adeno-Associated Virus include all of the following EXCEPT: A) Has a relatively small genome (4.8 kB) therefore large genes cannot be introduced into the virus and maintain viral activity B) Small virus not known to cause any human disease C) Infects both dividing and non-dividing cells D) Integrates DNA into a specific site on human chromosome 19
A is a disadvantage
Mechanical Transfection Method Characteristics: A) small gold particles coated with DNA B) Tissue must be accessible C) High intergration of DNA D) Good for short term gene expression.
C (low)
Which of the following is FALSE? Receptor-mediated Endocytosis:
A)
T/F Cationic Polymer Complexes allow the plasmid to move into the nucleus.
False, they help the plasmid move into the cell ans then are degraded.
Which of the following is NOT a CPC polymer?
A)
Which of the following I has a trivalent headgroup? Making it more positively charged. A) Lipofectamine B) DOTAP C) DC-Chol D) Glycofectamine
