27 Questions
What is the primary function of cell-penetrating peptides (CPPs) in DNA transformation?
Assisting in the penetration of nucleic acids into cells
What characteristic distinguishes CPPs from other cell-penetrating compounds?
Small peptide size
What is the potential benefit of using protein vehicles for DNA transformation in algae?
Enhancing transformation efforts
What type of interaction enables protein molecules to assist in DNA penetration into cells?
Non-covalent interactions
Why are CPPs commonly used for DNA transformation?
Due to their ability to facilitate DNA penetration into cells
What is the main limitation of conventional chalcone synthesis methods?
Prolonged reaction times, harsh conditions, toxic reagents, low selectivity, and low yields.
What is the novel approach presented to overcome the limitations of conventional chalcone synthesis methods?
Harnessing the power of genetic engineering in microorganisms through the incorporation of the chalcone synthase (CHS) gene into the microalga Chlamydomonas reinhardtii.
How is the genetic transformation facilitated in C. reinhardtii protoplasts?
Through biolistic gun, enabling the transfer of the CHS gene alongside a selectable Kanamycin resistance marker.
What is the primary application of the biolistic gun in agriculture?
Revolutionizing plant genetic modification, particularly in crops resistant to conventional methods, enhancing traits like pest resistance and yield.
How does the biolistic gun contribute to neuroscience research?
By enabling precise gene delivery into neuronal cells, advancing research on brain function and disorders.
What is the purpose of the biolistic gun technique in this project?
To transfer the CHS gene into C. reinhardtii for efficient chalcone production.
What is the significance of enhancing stress conditions in the microalga C. reinhardtii?
To induce the phenylpropanoid pathway, thereby increasing the yield of chalcones.
What is the role of the CHS gene in chalcone synthesis?
It is responsible for the production of chalcones.
What are the advantages of using biolistic particle delivery for gene transformation?
The advantages include being easy to use, fast, and versatile; not limited to host specificity or certain species; possibility of transient and stable expression; and not requiring large amounts of nucleic acids and cells.
Why is Chlamydomonas reinhardtii (C.Reinhardtii) an ideal organism for biolistic particle delivery?
C.Reinhardtii is ideal due to its ability to rapidly reproduce, has simple genetics, and the ability to photosynthesize.
What is the goal of using biolistic particle delivery in the project described?
The goal is to unlock the potential of C.Reinhardtii as a bio factory for producing valuable secondary metabolites like chalcone.
What is the significance of biolistic particle delivery in avoiding the use of viruses or chemicals?
Biolistic particle delivery avoids using viruses or chemicals which can be potentially toxic.
What is the benefit of using biolistic particle delivery for gene silencing?
Large DNA fragments may be transferred or small interfering RNAs during gene silencing.
What is the product that can be biosynthesized by C.Reinhardtii cells after the transfer of the chalcone synthase (CHS) gene?
Chalcone.
What is the basic principle behind the biolistic method of gene transfer?
Coating DNA onto microscopic carriers and shooting them into target cells at high speed.
What is the advantage of using the biolistic method for gene transfer in plants?
It can get DNA through thick tissues, making it useful for transforming plants that are hard to reach.
What is the characteristic molecular structure of chalcones?
Two aromatic rings connected by a three-carbon, α, β-unsaturated carbonyl system.
What is the role of chalcones in the plant kingdom?
They serve as precursors to several biologically active secondary metabolites.
What is the primary application of the biolistic method in genetic engineering?
Modulating metabolic pathways in organisms like Chlamydomonas reinhardtii.
What is the name of the device used in the biolistic method of gene transfer?
Gene gun.
What type of compounds are chalcones?
Polyphenolic compounds.
What is the current state of chalcone production?
Several synthetic methods are used for chalcone production.
Study Notes
Cell-Penetrating Peptides
- Protein vehicles show promise for enhancing DNA transformation in algae by facilitating the penetration of nucleic acids into cells and their integration into the genome.
- Protein molecules can assist in DNA transformation through non-covalent interactions with DNA structures.
- Cell-penetrating peptides (CPPs) are small peptides commonly used for DNA transformation.
- CPPs are the most widely used among different cell-penetrating compounds.
Chalcone Synthesis Limitations
- Prolonged reaction times, harsh conditions, toxic reagents, low selectivity, and low yields are drawbacks of conventional chalcone synthesis methods.
Novel Approach to Chalcone Synthesis
- Involves incorporating the chalcone synthase (CHS) gene from Petunia hybrida into microalga Chlamydomonas reinhardtii using genetic engineering.
- Enhancing stress conditions to induce the phenylpropanoid pathway increases chalcone yield.
- Genetic transformation is facilitated through biolistic gun in C. reinhardtii protoplasts.
Applications of Biolistic Gun
- Revolutionizes plant genetic modification, particularly in crops resistant to conventional methods, enhancing traits like pest resistance and yield.
- Enables precise gene delivery into neuronal cells, advancing research on brain function and disorders.
- Facilitates transient gene expression studies, aiding in understanding gene functions and cellular processes.
- Delivers therapeutic genes for gene therapy and DNA vaccination, and studies cancer biology.
- Allows tissue-specific gene delivery, contributing to research on organ development and diseases.
- Manipulates gene expression to unravel gene functions and regulatory networks in functional genomics.
CHS Gene Transfer through Biolistic Method
- Transfers CHS gene into C. reinhardtii for efficient chalcone production.
- Particle bombardment facilitates gene transfer, enabling the microalgae to biosynthesize chalcone.
Advantages of Biolistic Method
- Easy to use, fast, and versatile.
- Not limited to host specificity or specific species.
- Enables transient and stable expression.
- Only requires small amounts of nucleic acids and few cells.
- Can transfect many cell types, including non-dividing cells and plants.
- Avoids using viruses or potentially toxic chemicals.
- Facilitates gene silencing with large DNA fragments or small interfering RNAs.
Learn about cell-penetrating peptides, small peptides that facilitate DNA penetration into cells and integration into the genome. Understand their role in DNA transformation, particularly in algae.
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