2 - BIOTECH TOOLS

• What is biotechnology?

  The application of biological systems, organisms, or derivatives to create products or processes for specific uses
• What is the purpose of the Polymerase Chain Reaction (PCR)?

  To amplify (make many copies of) a specific DNA segment
• How does PCR mimic DNA replication in vitro?

  It uses a heat-stable DNA polymerase, such as Taq polymerase, to repeatedly copy the target sequence
• What are the main steps in the Polymerase Chain Reaction (PCR)?

  1. Denaturation: DNA is heated to separate the strands
  2. Annealing: Short DNA sequences (primers) bind to the target DNA
  3. Extension: DNA polymerase synthesizes new DNA strands complementary to the templates
  4. Repeat steps 1-3 for multiple cycles to exponentially amplify the target DNA
• What is recombinant DNA technology?

  The process of combining DNA from two or more different sources into one continuous DNA molecule
• What are the main steps in recombinant DNA technology?

  1. Isolate the gene of interest (often using PCR or restriction enzymes)
  2. Insert the gene into a plasmid (vector)
  3. Introduce the recombinant plasmid into a host cell (transformation)
  4. Select for host cells carrying the recombinant DNA
  5. Express the gene's protein product in the host cell
• What is molecular cloning?

  The process of producing identical copies of a DNA fragment by inserting it into a vector and replicating in a host
• What is a plasmid?

  A circle of double-stranded DNA that is separate from the chromosomes, found in bacteria and protozoa
• What is reproductive cloning?

  The process of producing genetically identical organisms
• What is the process of reproductive cloning?

  • Parthenogenesis: An embryo grows and develops without fertilization
  • The haploid nucleus of an egg cell is replaced with a diploid nucleus from a somatic cell of the same species
  • This creates a zygote that is genetically identical to the donor
• What is CRISPR-Cas9 genome editing?

  A technology that can precisely cut and modify DNA sequences inside living cells
• How does CRISPR-Cas9 genome editing work?

  1. Guide RNA (gRNA) directs the Cas9 enzyme to a specific DNA sequence
  2. Cas9 acts like molecular scissors to cut the DNA
  3. Cell's repair mechanisms:

  • Non-Homologous End Joining (NHEJ): Often introduces small insertions/deletions (gene knockout)
  • Homology-Directed Repair (HDR): Can insert specific DNA changes if a repair template is provided
• How was CRISPR originally discovered?

  As a bacterial defense system against viruses
• How does the CRISPR bacterial defense system work?

  • When bacteria kill a viral invader, proteins scoop up the remains of the virus's genetic code and cut it into tiny bits
  • The fragments are stored in CRISPR spaces in the bacterium's genome
  • The bacterium then uses this genetic information to fend off future attacks from the same viral invader
• What are the key differences between molecular cloning and reproductive cloning?

  Molecular Cloning:

  • Produces identical copies of a DNA fragment
  • Foundation for recombinant DNA work

  
  

  Reproductive Cloning:

  • Produces genetically identical organisms
  • Example: Dolly the sheep (via somatic cell nuclear transfer)
• How does the CRISPR-Cas9 system differ from other genome editing tools?

  • CRISPR uses a guide RNA to direct the Cas9 enzyme to a specific DNA sequence
  • Other tools, like zinc finger nucleases and TALENs, use engineered proteins to target DNA
  • CRISPR is simpler, more efficient, and more precise than previous genome editing methods
• What are the key differences between molecular cloning and reproductive cloning?

  Molecular Cloning:

  • Produces identical copies of a DNA fragment
  • Foundation for recombinant DNA work

  
  

  Reproductive Cloning:

  • Produces genetically identical organisms
  • Example: Dolly the sheep (via somatic cell nuclear transfer)
• What is the purpose of the CRISPR-Cas9 system?

  To precisely cut and modify DNA sequences inside living cells
• What are the key steps in the CRISPR-Cas9 genome editing process?

  1. Guide RNA (gRNA) directs Cas9 to a specific DNA sequence
  2. Cas9 enzyme acts like molecular scissors to cut the DNA
  3. Cell's repair mechanisms:

  • Non-Homologous End Joining (NHEJ): Often introduces small insertions/deletions (gene knockout)
  • Homology-Directed Repair (HDR): Can insert specific DNA changes if a repair template is provided
• What is the origin of the CRISPR-Cas9 system?

  It was originally discovered as a bacterial defense system against viruses
• How does the CRISPR-Cas9 system differ from other genome editing tools like zinc finger nucleases and TALENs?

  • CRISPR uses a guide RNA to direct the Cas9 enzyme to a specific DNA sequence
  • Other tools use engineered proteins to target DNA
  • CRISPR is simpler, more efficient, and more precise than previous genome editing methods
• Why is the CRISPR-Cas9 system considered a revolutionary genome editing tool?

  It is simpler, more efficient, and more precise than previous genome editing methods
• What are the key differences between molecular cloning and reproductive cloning?

  Molecular Cloning:

  • Produces identical copies of a DNA fragment
  • Foundation for recombinant DNA work

  
  

  Reproductive Cloning:

  • Produces genetically identical organisms
  • Example: Dolly the sheep (via somatic cell nuclear transfer)
• What is the purpose of recombinant DNA technology?

  To combine DNA from two or more different sources into one continuous DNA molecule
• What are the main steps in recombinant DNA technology?

  1. Isolate the gene of interest (often using PCR or restriction enzymes)
  2. Insert the gene into a plasmid (vector)
  3. Introduce the recombinant plasmid into a host cell (transformation)
  4. Select for host cells carrying the recombinant DNA
  5. Express the gene's protein product in the host cell
• What is the purpose of molecular cloning?

  To produce identical copies of a DNA fragment by inserting it into a vector and replicating in a host
• How does the CRISPR bacterial defense system work?

  • When bacteria kill a viral invader, proteins scoop up the remains of the virus's genetic code and cut it into tiny bits
  • The fragments are stored in CRISPR spaces in the bacterium's genome
  • The bacterium then uses this genetic information to fend off future attacks from the same viral invader
• What is the difference between molecular cloning and reproductive cloning?

  Molecular cloning produces identical copies of a DNA fragment, while reproductive cloning produces genetically identical organisms
• How does the CRISPR-Cas9 system work to edit genomes?

  1. Guide RNA (gRNA) directs the Cas9 enzyme to a specific DNA sequence
  2. Cas9 acts like molecular scissors to cut the DNA
  3. Cell's repair mechanisms:

  • Non-Homologous End Joining (NHEJ): Often introduces small insertions/deletions (gene knockout)
  • Homology-Directed Repair (HDR): Can insert specific DNA changes if a repair template is provided
• How does the CRISPR-Cas9 system differ from other genome editing tools like zinc finger nucleases and TALENs?

  • CRISPR uses a guide RNA to direct the Cas9 enzyme to a specific DNA sequence
  • Other tools use engineered proteins to target DNA
  • CRISPR is simpler, more efficient, and more precise than previous genome editing methods
• What is the key purpose of the CRISPR-Cas9 system?

  To precisely cut and modify DNA sequences inside living cells