Gene Therapy and Cell Culture Techniques

This set of flashcards is designed to help students understand key concepts related to gene therapy, cell culture techniques, and stem cell research, based on the lecture notes provided.

What is gene therapy?

A technique that modifies a person's genes to treat or cure disease.

What does 'in vivo' mean in gene therapy?

Refers to gene therapy conducted within a living organism.

What does 'ex vivo' mean?

Refers to gene therapy conducted on cells outside of the body, which are then returned to the patient.

What are somatic mutations?

Mutations that occur in somatic cells and are not inherited.

What are germline mutations?

Mutations that occur in gametes and can be passed to offspring.

What are autologous cells?

Cells derived from the same individual being treated.

What are allogenic cells?

Cells derived from a genetically different individual of the same species.

What is an episome?

A genetic structure in cells that is similar to a plasmid and can replicate independently.

What is a vector in gene therapy?

A carrier used to deliver genes into cells.

What is transduction?

The process of introducing foreign DNA into a cell using a vector.

What is gene augmentation therapy (GAT)?

A gene therapy strategy used to add functional copies of a gene to compensate for mutations.

What is the targeted killing of specific cells in gene therapy?

A strategy aimed at selectively destroying diseased cells.

When is a prodrug used in targeted killing?

A non-active drug that is converted into an active form within the body.

What is a suicide/toxin gene in gene therapy?

A gene that, when activated, causes the death of a targeted cell.

What is targeted mutation correction?

Gene therapy aimed at correcting specific mutations in DNA.

What are the two levels of targeted mutation correction?

Single-nucleotide correction and larger gene editing.

What does targeted inhibition of gene expression do?

Reduces or eliminates the expression of certain genes.

What cells do current gene therapy strategies typically target?

Somatic cells.

What are the two routes of administration for gene therapy?

Ex-vivo and in-vivo.

What is the general process for ex-vivo gene therapy?

Cells are removed from the patient, genetically modified, and then returned.

What is the general process for in-vivo gene therapy?

Gene therapy is directly administered into the body, targeting specific cells.

What is the significance of gene expression levels and DNA fragment size?

They affect the efficacy and safety of gene therapy.

What are the two methods of gene integration?

Random integration and targeted integration.

What are the advantages of viral vectors in gene therapy?

High efficiency of gene delivery and stable integration into the host genome.

What are the disadvantages of viral vectors?

Immunogenic responses and potential for insertional mutagenesis.

What are three types of non-viral vectors?

Liposomes, nanoparticles, and plasmid DNA.

What is the ideal disorder makeup for gene therapy?

Monogenic disorders that have clear genetic causes.

What gene therapy strategy is often used for inherited disorders?

Gene augmentation therapy.

What are three gene therapy options for pets?

Gene augmentation, gene editing, and somatic gene therapy.

What is tissue culture?

A technique for maintaining and growing cells in controlled conditions.

What is organ culture?

A method of maintaining whole organs in vitro to study their functions.

What is cell culture (CC)?

A technique used to grow cells under controlled conditions.

What is primary cell culture?

Cells that are directly taken from living tissue and cultured.

What is continuous cell culture?

Cells that can divide indefinitely in culture.

What are infinite cell cultures?

Cell lines that have been transformed to proliferate indefinitely.

What are finite cell cultures?

Cell lines that have a limited number of divisions before they cease to grow.

What is subculturing (passage) of cells?

The process of transferring cells to new culture vessels to allow further growth.

What is transformation in cell culture?

The process by which normal cells become cancerous.

What is transfection?

The introduction of foreign DNA into cells.

What is a solid phase in cell culture?

Refers to the culture medium designed to support cell attachment and growth.

What is a liquid phase in cell culture?

The culture medium that provides nourishment and environment for cells.

What does gaseous phase refer to in cell culture?

The atmosphere surrounding cultured cells that supplies oxygen and removes carbon dioxide.

What factors affect cell culture success?

Nutrient supply, pH, temperature, contamination, and cell line characteristics.

What is Hayflick's phenomenon?

The limited number of times a normal somatic cell can divide before cell division stops.

What is cryopreservation?

The process of freezing cells to preserve them for future use.

Why is cryopreservation important for cell culture?

It allows long-term storage of cells without losing viability.

What is a common dilemma with cell culture?

Maintaining cell viability and function over extended periods.

What are stem cells (SC)?

Cells that can differentiate into various types of cells and can self-renew.

What does self-renewable mean in stem cells?

The ability of stem cells to divide and produce more stem cells.

What is SC repair?

The capacity of stem cells to fix damaged tissues.

What does totipotent mean?

Stem cells that can differentiate into any cell type, including placental cells.

What does pluripotent mean?

Stem cells that can differentiate into nearly any cell type, excluding placental cells.

What does multipotent mean?

Stem cells that can develop into multiple but limited cell types.

What is a blastocyst?

An early stage of embryonic development consisting of an inner cell mass.

What is a feeder layer?

A layer of cells that provides essential support for another cell type.

What is differentiation in stem cells?

The process whereby a stem cell develops into a more specialized cell type.

What are embryoid bodies?

3D aggregates of stem cells that can differentiate into various cell types.

What are somatic stem cells?

Stem cells found in adult tissues that can differentiate into specialized cells.

What are hematopoietic stem cells?

Stem cells responsible for producing blood cells.

What are bone marrow stromal stem cells?

Stem cells found in bone marrow involved in hematopoiesis.

What is transdifferentiation?

The process by which one specialized cell type transforms into another.

What is reprogramming in stem cells?

The process of converting differentiated cells back into a stem cell-like state.

What are induced pluripotent stem cells (iPSCs)?

Stem cells created by reprogramming somatic cells to an embryonic stem cell-like state.

What are the three kinds of stem cell lines?

Embryonic stem cells, embryonic germ cells, and adult stem cells.

What characterizes embryonic stem cells?

Derived from embryos and are pluripotent, capable of becoming any cell type.

What are embryonic germ cells?

Stem cells derived from the early gonads of embryos.

What characterizes adult stem cells?

Found in adult tissues, multipotent, and involved in tissue maintenance.

What are two fundamental properties of stem cells?

Self-renewal and differentiation.

What are the two types of differentiation signals?

Internal signals from genes and external signals from the microenvironment.

What are three external signals that affect differentiation?

Chemical composition of the culture medium, physical contact with neighboring cells, and surrounding molecules.

What is the source of embryonic stem cells in research?

Derived from human embryos, specifically inner cell mass of the blastocyst.

What is the general process for stem cell cultures?

Isolating, expanding, and differentiating stem cells under controlled conditions.

What is a common coating type used in stem cell culture dishes?

Extracellular matrix proteins or synthetic coatings to provide attachment.

What is a concern with this coating type?

Potential for variability and inconsistency in cell attachment and growth.

What is the difference between transdifferentiation and reprogramming of adult stem cells?

Transdifferentiation refers to directly converting one type of cell into another, while reprogramming refers to converting specific cells back into stem cells.

How are iPSCs created?

By introducing specific genes associated with pluripotency into somatic cells.

What are three general categories of stem cell research applications?

Regenerative medicine, developmental biology, and disease modeling.

What are four unknowns in stem cell research?

Long-term effects, potential for tumor formation, mechanisms of differentiation, and the best sources of cells.

What are two challenges to stem cell research?

Ethical considerations and technological limitations.

What is the significance of the CLN5 gene?

It is implicated in Batten disease, a neurodegenerative disorder.

What is the primary method for analyzing data from gene therapy studies?

Statistical analysis for significance and efficacy.

What types of vectors are commonly used in gene therapy?

Viral vectors (e.g., AAV, lentiviruses) and non-viral methods (e.g., plasmids, liposomes).

What is the rationale behind the use of AAV vectors in gene therapy?

AAV vectors are known for their safety, ability to transduce non-dividing cells, and long-term expression.

What is the advantage of using sheep models in gene therapy research?

They closely mimic human diseases and provide a larger organ structure for studies compared to rodent models.

What role does the blood-brain barrier play in gene therapy?

It limits the types of molecules that can enter the brain, complicating the delivery of therapies.

How does CRISPR technology relate to gene therapy?

CRISPR allows for precise editing of genes, enabling corrections of mutations directly in target cells.

What are the ethical considerations in gene therapy research?

They include potential genetic alterations, consent, and long-term societal impacts.

What is gene editing?

Techniques used to modify the DNA of an organism, including CRISPR and TALENs.

What are potential applications of gene therapy?

Treating inherited genetic disorders, certain cancers, and viral infections.

What is an investigational new drug (IND) application?

A request to the FDA to begin testing a new drug or treatment in humans.

What role do vectors play in gene transfer?

They facilitate the delivery of genetic material into target cells.

How does cell culture technology advance gene therapy research?

It provides a controlled environment for studying cellular responses to therapies.

What is the significance of animal models in gene therapy?

They help to evaluate the efficacy and safety of therapies before human trials.

What do clinical trials for gene therapy assess?

Safety, effectiveness, and optimal dosing of the therapeutic intervention.

What is the importance of the chicken beta actin (CBh) promoter?

It is used to drive the expression of transgenes in various cell types.

What does 'transduced' refer to?

Cells that have taken up the genetic material through transduction.

What is the aim of the therapeutic window in gene therapy?

To determine the optimal dosages that maximize efficacy while minimizing adverse effects.

What is a significant challenge in developing effective gene therapies?

Ensuring long-term safety and efficacy without adverse immune responses.

What is the role of the Institutional Review Board (IRB) in gene therapy research?

To review research proposals to ensure ethical and safe study practices involving human participants.

What is the difference between random and targeted integration methods?

Random integration inserts genes at random locations in the genome, while targeted integration places genes at specific sites.