Recombinant DNA Technology and Gene Cloning
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Questions and Answers

What is the primary purpose of microinjecting foreign genes into eggs?

  • To increase the size of the fertilized eggs
  • To improve the taste of the animal
  • To fertilize the eggs more effectively
  • To produce transgenic offspring (correct)

    • Which gene is incorporated into fish to promote larger size?

  • Atlantic salmon gene
  • Bovine growth hormone gene
  • Ocean Pout gene promoter
  • Chinook salmon growth hormone gene (correct)

    • What is gene pharming?

  • Using farm animals for genetic research
  • Creating genetically modified crops
  • Farming animals for meat consumption
  • Using transgenic animals to produce pharmaceuticals (correct)

    • What notable characteristic does the AquAdvantage salmon have?

    <p>It reaches adult size three times faster than wild salmon</p> Signup and view all the answers

    What is one method for introducing DNA into eggs?

    <p>Hand microinjection</p> Signup and view all the answers

    What is the challenge associated with cloning animals?

    <p>Low success rate of producing viable embryos</p> Signup and view all the answers

    Which animal was the first cloned mammal?

    <p>Dolly the sheep</p> Signup and view all the answers

    How are proteins harvested in gene pharming?

    <p>From animal's milk</p> Signup and view all the answers

    What is the primary function of the Cas9 enzyme in the CRISPR system?

    <p>To break both strands of DNA in specific sequences</p> Signup and view all the answers

    How does CRISPR identify the genomic DNA of an invading virus?

    <p>Using a guide RNA that base-pairs with the DNA sequence</p> Signup and view all the answers

    What can be a potential application of CRISPR technology?

    <p>Developing new rapid tests for viruses like SARS-CoV-2</p> Signup and view all the answers

    What term is used to describe organisms that have foreign genes inserted into their genomes?

    <p>Transgenic organisms</p> Signup and view all the answers

    What is a common method for producing transgenic bacteria?

    <p>Recombinant DNA technology</p> Signup and view all the answers

    What do genetically modified organisms (GMOs) primarily produce?

    <p>Biotechnology products</p> Signup and view all the answers

    What is the role of RNA in the CRISPR system?

    <p>To carry instructions to the Cas9 enzyme</p> Signup and view all the answers

    What is one of the ongoing goals for scientists working with the CRISPR system?

    <p>To make the system more efficient and develop new applications</p> Signup and view all the answers

    What is the primary role of restriction enzymes in recombinant DNA technology?

    <p>To cleave vector DNA at specific sequences</p> Signup and view all the answers

    What are 'sticky ends' in the context of recombinant DNA?

    <p>Single-stranded, complementary ends of DNA molecules</p> Signup and view all the answers

    Which of the following best describes a vector in recombinant DNA technology?

    <p>A piece of DNA that can carry foreign DNA into a host cell</p> Signup and view all the answers

    What function does DNA ligase serve in recombinant DNA technology?

    <p>It facilitates the binding of foreign DNA to the plasmid</p> Signup and view all the answers

    Which statement about plasmids is true?

    <p>They are used as vectors for cloning foreign DNA</p> Signup and view all the answers

    How do restriction enzymes benefit bacteria?

    <p>By cutting up viral DNA to prevent infection</p> Signup and view all the answers

    What must be true for foreign DNA to be successfully inserted into a vector?

    <p>Complementary ends must be created using the same restriction enzyme</p> Signup and view all the answers

    What is the significance of having hundreds of types of restriction enzymes in bacteria?

    <p>They enable bacteria to cut DNA at multiple specific sites</p> Signup and view all the answers

    What is the primary purpose of gene therapy?

    <p>To insert genetic material into human cells for treating disorders</p> Signup and view all the answers

    What is a key component in the ex vivo gene therapy method for SCID treatment?

    <p>Bone marrow stem cells</p> Signup and view all the answers

    Which component is commonly used to transfer normal genes into the body in gene therapy?

    <p>Viruses or liposomes</p> Signup and view all the answers

    What is the first step in the ex vivo gene therapy process for treating SCID?

    <p>Removing bone marrow stem cells from the patient</p> Signup and view all the answers

    What characteristic do the female clones of transgenic animals share with the original donor?

    <p>Same product in their milk</p> Signup and view all the answers

    How are viruses utilized in gene therapy?

    <p>They are genetically modified to become safe carriers of normal genes.</p> Signup and view all the answers

    What is the role of liposomes in gene therapy?

    <p>To transport and deliver normal genes into cells</p> Signup and view all the answers

    Which condition is specifically treated using the ex vivo method of gene therapy?

    <p>SCID (severe combined immunodeficiency)</p> Signup and view all the answers

    What is the primary purpose of ex vivo gene therapy in the treatment of familial hypercholesterolemia?

    <p>To add a normal gene for cholesterol removal</p> Signup and view all the answers

    In cystic fibrosis treatment, which method is used to deliver the gene needed for therapy?

    <p>Direct delivery via liposomes or adenovirus vector</p> Signup and view all the answers

    Which gene is often targeted in cancer gene therapy to make tumor cells more sensitive to chemotherapy?

    <p>p53</p> Signup and view all the answers

    RNA interference (RNAi) primarily functions by targeting which type of RNA?

    <p>Messenger RNA</p> Signup and view all the answers

    What was the original discovery context for RNA interference?

    <p>In worms as a protective mechanism against viruses</p> Signup and view all the answers

    In the context of gene therapy, what is the significance of using adenovirus vectors?

    <p>They efficiently deliver genes to respiratory tract cells</p> Signup and view all the answers

    What is one of the main challenges faced in in vivo gene therapy for cystic fibrosis?

    <p>Limited success in gene delivery</p> Signup and view all the answers

    What is the mechanism by which RNA interference silences specific alleles?

    <p>By producing double-stranded RNA to be degraded</p> Signup and view all the answers

    Study Notes

    Recombinant DNA (rDNA) Technology

    • rDNA is DNA that contains genetic material from two or more different sources.
    • Vectors carry foreign DNA into a cell.
    • Plasmids are accessory rings of DNA found in bacteria and commonly used as vectors.
    • Plasmids are not part of the bacterial chromosome.

    Cloning a Human Gene

    • Restriction enzymes cleave vector DNA at specific sites.
    • They act as molecular scissors for cutting DNA in precise sequences.
    • DNA ligase seals foreign DNA into the opening in the vector DNA created by the restriction enzyme.

    Restriction Enzymes

    • Restriction enzymes were first discovered in prokaryotes as an immune defense against invading viruses.
    • The CRISPR system involves an endonuclease enzyme called Cas9.
    • Cas9 recognizes specific nucleotide sequences in the genomic DNA of invading viruses, and breaks both DNA strands, inactivating the virus.
    • CRISPR utilizes a guide RNA molecule that complements the genomic DNA sequence.

    Genome Editing

    • Scientists can study a gene's role in cells after Cas9 inactivates the gene.
    • Cas9 allows insertion of new nucleotides at specific DNA locations.
    • CRISPR can target specific nucleotide sequences for editing in almost any organism.

    CRISPR and Genome Editing Applications

    • CRISPR applications include treating diseases such as sickle-cell disease and cancer.
    • CRISPR can also be used for rapid virus testing, such as for SARS-CoV-2.

    Biotechnology Products

    • Organisms with an inserted foreign gene are called transgenic organisms.
    • Transgenic bacteria, plants, and animals are often called genetically modified organisms (GMOs).
    • The products produced by transgenic organisms are called biotechnology products.

    Genetically Modified Bacteria

    • Transgenic bacteria are produced using recombinant DNA technology.
    • They are grown in large vats called bioreactors.
    • Bacteria express the cloned gene.
    • The gene product is collected from the media.

    Genetically Modified Animals

    • The AquAdvantage salmon is a transgenic fish 99.9% Atlantic salmon but with gene products from two other fish:
      • Growth hormone gene from the Chinook salmon for faster growth.
      • Growth hormone gene promoter from the Ocean Pout to keep the growth hormone gene active.
    • The gene products are grown on bacterial plasmids, isolated, then mixed with fertilized Atlantic salmon eggs.
    • AquAdvantage salmon reach adult size 3 times faster than wild salmon using less food.

    Producing Transgenic Animals

    • Gene pharming uses transgenic farm animals to produce pharmaceuticals.
    • Therapeutic and diagnostic protein genes are incorporated into an animal's DNA.
    • Proteins can be harvested from the animal's milk.
    • Potential for producing drugs for cystic fibrosis, cancer, and blood diseases.

    Cloning Transgenic Animals

    • Scottish scientists cloned the sheep Dolly in 1997.
    • Since then, calves, goats, pigs, rabbits, and cats have been cloned.
    • The process is difficult and has a low success rate.
    • Approximately 1-2 viable embryos are produced per 100 attempts.

    Cloning Transgenic Animals Process

    • Donor eggs are microinjected with nuclei from one transgenic animal.
    • Eggs are coaxed to begin development in vitro.
    • Development continues in host females until clones are born.
    • Female clones produce the same product in their milk as the original transgenic animal.

    Gene Therapy

    • Gene therapy involves inserting genetic material into human cells to treat genetic disorders and other illnesses, like cardiovascular disease and cancer.
    • Various methods of gene transfer are used.
    • Genetically modified viruses can introduce a normal gene into the body.
    • Liposomes (microscopic globule of lipids) can introduce normal genes.
    • Direct gene injection into a specific region of the body is also possible.

    Ex Vivo Gene Therapy

    • Ex vivo method for treating SCID (severe combined immunodeficiency).
    • Used for children lacking the enzyme ADA (adenosine deaminase) involved in T and B cell maturation.
    • SCID patients are prone to constant infections and may die without treatment.

    Ex Vivo Gene Therapy Treatment Steps:

    • Bone marrow stem cells are removed from the patient's body.
    • Cells are infected with a virus that carries the normal gene for the enzyme ADA.
    • Cells are returned to the patient with the hope that they will divide, expressing the normal gene for ADA.

    Ex Vivo Gene Therapy for Familial Hypercholesterolemia

    • Liver cells lack a receptor protein for removing cholesterol from the blood in patients with familial hypercholesterolemia.
    • High blood cholesterol levels put individuals at risk for heart attacks at a young age.
    • A portion of the liver is surgically excised and infected with a virus carrying the normal gene for the receptor.
    • The liver portion is then returned to the patient.

    In Vivo Gene Therapy

    • In vivo gene therapy involves directly delivering a gene to the body.
    • Cystic fibrosis patients lack the gene coding for the chloride transporter membrane protein, leading to thick mucus accumulation in the lungs and respiratory tract infections.
    • Treatment involves spraying the gene needed to cure cystic fibrosis into the nose or delivering it to the lower respiratory tract with an adenovirus vector or liposomes.
    • Limited success has been achieved so far.

    In Vivo Gene Therapy for Cancer Treatment

    • In vivo gene therapy is increasingly used in cancer treatment.
    • It aims to make healthy cells more tolerant of chemotherapy and tumor cells more sensitive to chemotherapy.
    • Researchers are investigating how to introduce the tumor suppressor gene p53 into cancer cells.

    RNA Interference

    • RNA interference (RNAi) uses RNA pieces to silence the expression of specific alleles.
    • These RNA pieces are complementary to the mRNA transcribed by a certain gene.
    • They bind to the target RNA in the cell, forming double-stranded RNA molecules, which are then broken down by enzymes.

    RNA Interference Discovery and Evolutionary Significance

    • RNA interference was first discovered in worms.
    • It is believed to have evolved in eukaryotic organisms as a defense mechanism against viruses.

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    Description

    Explore the fascinating world of recombinant DNA technology and gene cloning through this quiz. Learn about the roles of restriction enzymes, plasmids, and CRISPR in gene manipulation. Test your understanding of the methods used to edit genomes and the significance of these innovations in biotechnology.

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