PART2(Postmidterm) Flashcards
is a process by which foreign DNA is transferred to host cells for applications such as genetic research or gene therapy.
Gene delivery
- a type of gene transfer that utilize bacterial or viral carriers such as Agrobacterium tumefaciens and Agrobacterium rhizogenes
Biological or Indirect
Two types of direct gene delivery method
Chemical method
Physical method
A direct method that include certain polymers and lipids,
Chemical methods
A direct method which utilize physical properties and forces to transport genetic material into cells.
Physical methods
- insertion of exogenous DNA into a prokaryotic cell
Transformation
- denotes gene delivery via a virus.
Transduction
- gene delivery via nonviral gene delivery method
transfection
delivers RNA into cells, enveloped structure and can deliver up to 8000 bases of ssRNA can yield permanent transduction
Retroviruses
a group of nonenveloped viruses that carry linear, dsDNA.
- occur naturally, infecting mucosal linings in humans and other mammals.
Adenoviruses
widespread; have the ability to infect a wide variety of plant species like wheat, maize, cotton, and more.
Geminiviruses
is a process of using Agrobacterium to transfer a gene of interest into the plant cells, generating transgenic plants.
Agrobacterium-mediated
- a soil pathogenic bacterium; causes crown-gall disease or hairy root disease in infected dicotyledonous plants.
Agrobacterium
A bacterium has a natural ability to transfer the so-called T-DNA in its Ti plasmid into plant cells
Agrobacterium tumefaciens
- facilitates the uptake of foreign DNA into the protoplasts by inducing transient pore formation in the plasma membrane, allowing the DNA to enter the cells.
PEG-mediated transformation
- Plant cells with cell walls removed, leaving only the plasma membrane intact. Protoplasts are used as the target cells for introducing foreign DNA, as their lack of cell walls makes it easier for the DNA to enter the cell.
Protoplasts
- acts as an osmotic stabilizer to help maintain the integrity and viability of the protoplasts during handling and transformation procedures.
Mannitol
is a gene that is attached to the regulatory sequence of another gene of interest
Reporter genes
are a type of marker genes that are used in the artificial selection of transformed cells; usually they are antibiotic or herbicide resistance genes
Selectable markers
A gene whose expression allows one to identify cells that have been transformed or transfected with a vector containing the marker gene
Selectable marker
A gene used to tag another gene or DNA sequence
Reporter marker
The DNA sequences can be inserted, deleted, modified or replaced, which is analogous to editing a manuscript.
Genome Editing
- This part is customizable and can be engineered to bind to any specific DNA sequence.
TALE DNA-Binding Domain
- This nuclease cuts the DNA when guided by the TALE domain.
Fokl Nuclease Domain
The most popular genome editing methods because of ease, precision and relatively lower cost
CRISPR/Cas
determines the order of the four chemical building blocks - called “bases” - that make up the DNA molecule.
DNA sequencing
A sequencing method that is also known as chemical sequencing because chemical reactions, rather than DNA and RNA amplification, are the basis of the method.
Maxam–Gilbert sequencing
A sequencing that is also known as the “chain termination method”, is a method for determining the nucleotide sequence of DNA
Sanger sequencing
is the process of generating a genetically identical copy of a cell or an organism
Cloning
DNA Cloning Application
- Cutting and Pasting DNA
- Bacterial transformation and selection
- Protein production
Two types of Gene delivery method
Biological or Indirect method
Direct method
Two types of Biological or Inderect gene delivery method
Agrobacterium-mediated transformation
Viral delivery
Two types of Direct Gene delivery method
Chemical method
Physical method
2 bacterium uses in Agrobacterium-mediated transformation
Agrobacterium tumefaciens
Agrobacterium rhizogenes
Viral delivery method
Retroviruses
Adenoviruses
Geminiviruses
Direct chemical methods
Liposomes
Polymers
Silicon carbide
Direct Physical methods
Gene gun
Electroporation
Ultrasonication
Microinjection
Macroinjection
Three main components in performing Agrobacterium-mediated transformation
T-binary system
Agrobacterium competent cells
Plants
Four methods in Genome Editing
Meganucleases
Zinc finger nucleases
TALEN
CRISPR/Cas
Two main components of TALEN
TALE DNA-Binding domain
Fokl nuclease domain
CRISPR two components
sgRNA
Cas endonuclease
CRISPR TWO STAGE GENOME EDITING
DNA damage generation
Host cell repair
First Generation DNA Sequencing Method
Maxam-Gilbert Sequencing
Sanger Sequencing
