Gene Therapy
What is gene therapy?
a treatment that aims to correct genetic defects or mutations that cause diseases
CRISPR was originally discovered in bacteria. What does it do in bacteria?
CRISPR is coupled with the Cas9 restriction enzyme to edit genes and is known as the CRISPR-Cas9 system.
Describe Somatic Cell Nuclear Transfer.
technique in which the nucleus of a somatic (body) cell is transferred to the cytoplasm of an enucleated egg
What does PCR stand for?
Polymerase Chain Reaction
What is an embryologist?
studies and works with embryos in human reproduction
What makes a good candidate for gene therapy?
-disease is caused by single gene mutation
-affected gene is understood
-target cells for therapy are accessible
- clear therapeutic benefit from correcting or replacing the gene
-disease had no conventional treatments
How can CRISPR be used by humans?
CRISPR can precisely modify a piece of DNA or its chemistry in the human body
What is the difference between reproductive cloning and therapeutic cloning?
Therapeutic cloning is the cloning of organ or tissues, reproductive cloning is the cloning of an entire person
Why is PCR performed?
Produces numerous copies of a specific segment of our DNA in a short period of time
What is tag polymerase?
enzyme that copies DNA from hot springs bacteria
what is a vector?
many different molecular tools or delivery systems
Has CRISPR ever been used in humans? When and where?
October 28, 2016 by a team from the Sichuan University in Chengdu
Who is Dolly?
Dolly is the first mammal created by Somatic Cell Nuclear Transfer
Describe the 3 steps of PCR including the temperature each is performed at
1)Denaturation- Heat to separate DNA strands (95C)
2)Annealing- cool to allow primers to bind (55C)
3) Extension- Heat slightly so that the Taq Polymerase extends the 3’ end of each primer (72C).
What is DNA primers?
Short single stranded DNA sequences complementary to beginning + end of gene you want to copy.
Factors to determine what vector to use?
-target cell type
-size of gene
-duration of expression
-immunogenicity
-safety
-production and purification
How is Cas9 used to specifically target a gene of interest?
-By designing a guide RNA that matches the target gene sequence,
-forming a Cas9 gRNA complex,
-binding to the target DNA,
-and utilizing the cells repair mechanisms to introduce desired genetic mechanisms.
What are some arguments against the use of cloning?
-ethical concerns
-identity and individuality
-health risks
-psychological impact
-biodiversity and genetic diversity
-exploitation and abuse
-religious and cultural objections
-legal and regulatory issues
What is a reproductive endocrinologist?
diagnose and treat endocrine disorders related to reproduction (infertility).
what are DNA nucleotides?
free floating nucleotides
what are current concerns/cons to using gene therapy?
-Expensive
-experimental/new
-potentially dangerous
-ethical issues
-possibility of infection
what is gene doping?
the use of gene therapy to modify genes that improve athletic performance.
Define in-vivo and ex-vivo
In vivo: The direct delivery of genes into the cells of a particular tissue. Ex vivo: This involves the transfer of genes in cultured cells which are then reintroduced into the patient.
what is an andrologist do?
knowledge and expertise to diagnose and treat male reproductive health issues.
what is a thermal cycler?
used to amplify segments of DNA via the polymerase chain reaction.