20 Questions
What is a characteristic of cyanobacteria that makes them amenable to DNA uptake?
They can natively take up DNA from their environment without manipulation
What is a challenge posed by eukaryotic algae in gene delivery methods?
Their requirement for aggressive delivery methods due to cell walls
What is a factor that influences the efficiency of gene delivery methods?
The target cellular compartment
What is a way in which eukaryotic algae differ from cyanobacteria in terms of DNA uptake?
Eukaryotic algae require more aggressive delivery methods
What is the importance of target cellular compartment in gene delivery methods?
It affects the efficiency of gene delivery
Why are certain cells more amenable to gene delivery methods?
Due to their ability to natively take up DNA from the environment
What is the primary application of microparticle bombardment in gene delivery?
Transformation of chloroplast genomes
How does the presence of cell walls affect transformation efficiency in microparticle bombardment?
It has no effect on transformation efficiency
What is the minimum reported transformation efficiency in C.reinhardtii using microparticle bombardment?
15 transformants per µg of DNA
What is the advantage of using smaller particles in microparticle bombardment?
It improves transformation efficiency
What is the name of the technique used to deliver proteins into target cells using microparticle bombardment?
Proteolistics
What is the reported efficiency of delivering Cas9–gRNA RNP into P.tricornutum using microparticle bombardment?
10⁻⁶ per 10⁶ cells mL⁻¹
What is the consequence of knocking out the PtUMPS and PtAPT genes in P.tricornutum using microparticle bombardment?
Resistance to 5-fluoroorotic acid and 2-fluoroadenine
What is the size range of proteins that can be delivered using microparticle bombardment?
24-68 kDa
What type of peptides have been demonstrated in non-algal hosts?
Cell penetrating peptides
What is a common application of metal-organic frameworks?
Gene delivery
What is a benefit of using liposomes in gene delivery?
Increased transformation efficiency
Which of the following is NOT a method of gene delivery mentioned in the text?
Viral vectors
What is the main goal of target cell recognition in gene delivery?
To specifically target cells for gene delivery
What is the focus of the review mentioned in the text?
Gene delivery methods in non-algal hosts
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).
This quiz covers the strategies and applications of cell penetrating peptides, polymers, metal-organic frameworks, and nanoparticles. It's ideal for biochemistry and nanotechnology students.
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