Podcast
Questions and Answers
What is a characteristic of cyanobacteria that makes them amenable to DNA uptake?
What is a characteristic of cyanobacteria that makes them amenable to DNA uptake?
- They have a cell wall that requires aggressive delivery methods
- They are eukaryotic organisms
- They can natively take up DNA from their environment without manipulation (correct)
- They require specialized organelles for DNA uptake
What is a challenge posed by eukaryotic algae in gene delivery methods?
What is a challenge posed by eukaryotic algae in gene delivery methods?
- Their lack of cell walls
- Their ability to recognize and reject foreign DNA
- Their requirement for aggressive delivery methods due to cell walls (correct)
- Their ability to natively take up DNA from the environment
What is a factor that influences the efficiency of gene delivery methods?
What is a factor that influences the efficiency of gene delivery methods?
- The type of DNA molecule being used
- The target cellular compartment (correct)
- The environmental conditions of the cell
- All of the above
What is a way in which eukaryotic algae differ from cyanobacteria in terms of DNA uptake?
What is a way in which eukaryotic algae differ from cyanobacteria in terms of DNA uptake?
What is the importance of target cellular compartment in gene delivery methods?
What is the importance of target cellular compartment in gene delivery methods?
Why are certain cells more amenable to gene delivery methods?
Why are certain cells more amenable to gene delivery methods?
What is the primary application of microparticle bombardment in gene delivery?
What is the primary application of microparticle bombardment in gene delivery?
How does the presence of cell walls affect transformation efficiency in microparticle bombardment?
How does the presence of cell walls affect transformation efficiency in microparticle bombardment?
What is the minimum reported transformation efficiency in C.reinhardtii using microparticle bombardment?
What is the minimum reported transformation efficiency in C.reinhardtii using microparticle bombardment?
What is the advantage of using smaller particles in microparticle bombardment?
What is the advantage of using smaller particles in microparticle bombardment?
What is the name of the technique used to deliver proteins into target cells using microparticle bombardment?
What is the name of the technique used to deliver proteins into target cells using microparticle bombardment?
What is the reported efficiency of delivering Cas9–gRNA RNP into P.tricornutum using microparticle bombardment?
What is the reported efficiency of delivering Cas9–gRNA RNP into P.tricornutum using microparticle bombardment?
What is the consequence of knocking out the PtUMPS and PtAPT genes in P.tricornutum using microparticle bombardment?
What is the consequence of knocking out the PtUMPS and PtAPT genes in P.tricornutum using microparticle bombardment?
What is the size range of proteins that can be delivered using microparticle bombardment?
What is the size range of proteins that can be delivered using microparticle bombardment?
What type of peptides have been demonstrated in non-algal hosts?
What type of peptides have been demonstrated in non-algal hosts?
What is a common application of metal-organic frameworks?
What is a common application of metal-organic frameworks?
What is a benefit of using liposomes in gene delivery?
What is a benefit of using liposomes in gene delivery?
Which of the following is NOT a method of gene delivery mentioned in the text?
Which of the following is NOT a method of gene delivery mentioned in the text?
What is the main goal of target cell recognition in gene delivery?
What is the main goal of target cell recognition in gene delivery?
What is the focus of the review mentioned in the text?
What is the focus of the review mentioned in the text?
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Study Notes
Cell Penetrating Peptides and Delivery Methods
- Strategies based on cell penetrating peptides (CPP), polymers, metal-organic frameworks, nanoparticles, and liposomes have been demonstrated in non-algal hosts.
Delivery to Cellular Compartments
- Methods can target specific cellular compartments (organellar or nuclear).
- Cyanobacteria can natively take up DNA from their environment without manipulation.
- Eukaryotic algae require more aggressive delivery methods due to their cell walls.
Transformation of Chloroplast Genomes
- Cell penetrating peptides can penetrate multiple membrane layers.
- This technique can transform circular plasmids into the nucleus of the red alga Porphyridium purpureum.
Microparticle Bombardment
- This method is unaffected by cell-wall presence but can disturb cell viability if many micro-projectiles are used.
- Efficiency of nuclear transformation in Chlamydomonas reinhardtii: 15 transformants per µg of DNA.
- Efficiencies in Dunaliella salina, Volvox carteri, and Phaeodactylum tricornutum: 2.5 × 10^(-5), 1.7 × 10^2, and 1.0 × 10^2 transformants per µg of DNA, respectively.
Proteolistics
- Microparticle bombardment can deliver proteins (24-68 kDa) into cells.
- This method has been used to deliver Cas9-gRNA ribonucleoprotein (RNP) into Phaeodactylum tricornutum, leading to gene knockout and resistance to 5-fluoroorotic acid (5-FOA) and 2-fluoroadenine (2-FA).
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