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DNA Technology

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Practical application of knowledge about DNA.

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Selective Breeding

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Artificial breeding to get a specific result/outcome.

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Recombinant DNA

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Genetic material cut by restriction enzymes and then spliced together from multiple sources (can be from two different sources, but doesn't have to).

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Transgenic

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Containing DNA from multiple species and the two different sources of information are added together.

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Examples of Transgenic organisms

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Yeasts and Bacteria

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Reverse Transcriptase

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Enzyme that uses RNA as a template to construct a DNA Molecule.

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Cloning Vector

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A self-replicating structure that carries DNA from cell to cell.

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Plasmid

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Small circle of double stranded DNA separates from a cell's chromosome.

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Restriction Enzymes

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Protein that cuts double-stranded DNA at a specific base sequence by "attacking" foreign genetic material at a specific base sequence (cut plasmid and add insulin).

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Transformation

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Introducing bacteria

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Sticky Ends

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Complementary to each other

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Electrophoresis

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Technique used to sort DNA fragments by size.

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Pseudogene

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A DNA sequence that is very similar to that of a gene; it is transcribed but its mRNA is not translated.

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Polymerase Chain Reaction (PCR)

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Biotechnology tool that rapidly produces millions of copies of a DNA sequence of interest (DNA Photo Copy Machine).

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What temperature does Polymerase Chain Reaction (PCR) need to be at and what enzyme can resist very hot temperatures?

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Needs to be at a specific temperature range or it will denature. TAQ Polymerase is an enzyme that can endure very hot temperatures.

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What are the steps of the enzyme TAQ Polymerase?

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1. Heat and strands separate 2. Primer 3. Complementary sides added 4. Cool

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DNA Profiling

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Biotechnology tool that uses DNA to detect genetic differences between individuals

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Short Tandem Repeats (STR)

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Short DNA sequences that vary in length among individuals in a population and occur in introns or non-coding regions

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Gel Electro

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Load DNA and Proteins, separate molecules by size, and the restriction enzyme helps cut the DNA

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Single Nucleotide Polymorphism (SNP) and when is it noticeable?

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One letter difference in two different strands and is noticeable when restriction enzymes are used because they help differentiate people.

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Stem Cells

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Any undifferentiated cell that can give rise to specialized cell types

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What happens when Stem Cells divide Mitotically ?

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Two daughter cells are yielded, one remains a stem cell- able to divide again

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Embryonic Stem Cells

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Stem cell that can give rise to all types of cells in the body (totipotent), very versatile because they form babies

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Adult Stem Cells

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Cell that can give rise to a limited subset of cells in the body (pluripotent), less abundant

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Cloning

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Asexual Reproduction

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Somatic Cell Nuclear Transfer

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Technique used to close a mammal from an adult cell

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Steps of a Somatic Cell Nuclear Transfer (5)

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1. Mammary glands consist of somatic cells, which are body cells that do not give rise to sperm or eggs 2. They then transferred this "donor" nucleus to the sheep's egg cell whose nucleus that also been removed 3. Embryo is formed by resulting mitotically divided cells 4. Researchers implanted in a surrogate mother's uterus 5. The embryo develops into the lamb

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Gene Therapy bullet points

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1. Provide new treatment options by adding healthy DNA to a person's cells 2. Use DNA to silence a gene whose activity is causing illness 3. Use a donor human cell 4. Would not able to pass gene onto children

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RNA Interference

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Prevent translation of mRNA from a cancer-related gene

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Karyotype (and example) - Genetic Test

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View, count, and sort the cell's chromosomes Example: Down Syndrome (Trisomy 21)

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Protein Production (Biochemical) (and example)- Genetic Test

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Measures the amount and quality of the protein present Example: Tax-Sachs

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Direct Test (and example)- Genetic Test

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Look at sequence (use DNA Probe) Example: Sickle-Cell

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PG10- Timing

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Examine embryos prior to inserting embryo into uterus (in vitro)

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Prenatal- Timing

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Examine fetus in womb; via amniocentesis and chronic villi sampling

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After Birth- Timing

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Carrier: Determine likelihood of offspring's traits Presymtomatic: Look for flawed gene Predicture: Will have a disorder or won't (PKO)

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Susceptibility

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Look for genes that have an increased likelihood of disease (no certainty)

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DNA Probe

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A single stranded sequence of nucleotides that is complementary to a particular region of DNA; short and labeled with radioactive isotope or tagged

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Preimplantation Genetic Diagnosis (PGD)

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In vitro fertilization-sperm fertilizes egg in dish which increases an identical embryo and removes one cell that undergoes PCR and shows the infected gene; very sensitive and room for human error

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Blastocyst (5 days old embryo)

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Shell of cells containing a lump of cells call the ICM

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Morula (3 days old embryo)

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A solid ball if cells resulting from division of a fertilized ovum

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Gastrula (18 days old embryo)

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An embryo at stage following the blastula, when it is a hollow-cup shaped structure having three layers if cells

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Differentiation

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Unspecialized stem cells give rise to specialized cells in response to internal and external chemical signs

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Potency

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The power of something to affect the mind or body

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Multipotency (potency)

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Only limited number of types of tissues; limited potential

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Pluripotent (potency)

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Almost all types of tissues; mostly potential

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Totipotent (potency)

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Differentiation into any cell type; total potential

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What are the three germ cell layers of a developing embryo? List them inside out.

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Entoderm: nervous system, skin Mesoderm: muscles, bones Ectoderm

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What is the ICM (Inner Cell Mass)

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Lump of cells are pluripotent which means they produce any of the 200 cell types found in the body, they can multiply in a dish.

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What are some of the potential uses of stem cells?

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Biological and Medical Research 1. Coax stem cells to divide to cure disorders 2. Reduce amount of need for laboratory animals

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What are some of the risks associated with stem cells?

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1. Patient's immune system could reject tissues 2. Could cause tissue or organ damage 3. Destruction of the embryo 4. Possible tumors

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How can scientists reprogram cells to become stem cells?

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Reprogramming a patients own cells by using fat, skin, and fibroblasts

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Describe the process of SCNT (Somatic Cell Nuclear Transfer).

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1. Somatic cell transferred to donor nucleus 2. Embryo forms 3. Implanted into Uterus 4. Embryo develops

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Describe the three types of cloning.

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Reproductive: Genetic Material of an adult somatic cell is transplanted into unfertilized egg that has no genetic material to make a child Therapeutic: Use of technology in the search for treatments and cures for diseases and disabilities Gene: Gene is copied when the DNA is extracted from one organism and another DNA is inserted in and copied

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What are the risks associated with cloning?

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1. Most die at an early age 2. Have abnormalities 3. Medical risks 4. Costs much money 5. Abnormal gene expression patterns 6. Telomeric differences

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Gene Therapy

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Technique for correcting defective genes responsible for disease development

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How does gene therapy work?

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1. Normal gene inserted to replace nonfunctional gene 2. Abnormal gene swapped with normal gene 2. Abnormal gene could be repaired through selective reverse mutation 2. Regulation of the abnormal gene could be altered (turned on/off)

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What criteria makes a disease a good candidate for gene therapy?

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- Mutation in one or more genes - "Single Gene" disorders ; mutation in a single gene - not having the mutated gene be "negative dominant"

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What is a vector? Give four examples.

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Vector is a molecular "delivery truck" usually made from a disabled virus Examples: Retroviruses, Adenoviruses, Aden-associated viruses, and Herpes Simplex viruses

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What are some challenges of gene therapy?

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-sometimes fatal side effects -not always successful

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What is the current status for gene therapy research?

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Currently, the FDA has not approved any human gene therapy product for sale. Although, their are experimental trials now.

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What ethical issues surround gene therapy?

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Gene therapy is not always effective but still can cause fatal side effects. Should gene therapy still be a solution?

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How did gene therapy help the boy in the bubble (SCID)?

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Scientists isolated the normal copy of the gene and packed it into a vector. The vector then was used to transport the gene into bone marrow cells. The bone marrow cells created the immune system.

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What are the steps in using a vector?

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1. Isolate normal DNA 2. Normal DNA packaged into Vector 3. Infect target (abnormal) cell 4. Vector "unloads" cargo (normal gene) 5. Proaction of Normal gene

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DNA microarrays contain ______________________. a) single stranded DNA b) complementary DNA c) genomic DNA d) double stranded DNA

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DNA microarrays contain a) single stranded DNA

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What was the source of the DNA for the cloning of the Banteng cow? a) an adult female's mammary cells b) an adult male's skin cells c) an adult female's uterine cells d) an adult male's sperm cells

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The source of the DNA for cloning the Bateng cow was b) an adult male's skin cells

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True or False? Although cloning endangered animals may increase the population size of Bateng cattle, it will not increase the genetic diversity of the species.

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True

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What is the original sequence of DNA? AGCTGCCATCTTG

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TCGACGGTAGAAC (Complementary)

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Which situation best represents how DNA sequencing, PCR, and gene probes might be used together? a) scientists might create a gene probe to find the correct DNA strand to sequence. Once it is sequenced, they might replicate to with PCR b) scientists might sequence a strand of DNA before replicating it with PCR. Once it is replicated, they can create a complementary gene probe c) scientists might replicate a strand of DNA using PCR before sequencing it. Once the sequence is known, they can produce a corresponding gene probe

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DNA sequencing, PCR, and gene probes might be used together when c) scientists might replicate a strand of DNA using PCR before sequencing it. Once the sequence is known, they can produce a corresponding gene probe

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True or False? Extracting a nucleus from a sheep cell and fusing it with the egg cell of a second sheep of the same species produces the first cell of a transgenic organism.

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False

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Which of the following statements are false regarding DNA? a) Doctors analyze DNA sequences to screen for diseases b) Scientists use DNA sequences to determine guilt in legal cases c) Proteins can be injected into cells to encode damaged DNA sequences d) Each person inherits DNA from two parents e) DNA encodes proteins f) Mitochondrial DNA is composed of the same four nucleotides as nuclear DNA

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false: c) Proteins can be injected into cells to encode damaged DNA sequences

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PGD

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- tests for genetic sequences in embryos - uses DNA probes

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Genetic Testing

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- tests for genetic sequences in developed individuals - uses DNA probes

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Gene Therapy (end)

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- replaces faulty genes - uses viruses

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Difference between stem cells and differentiated cells.

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stem cells: unspecialized, more common early in development than late in development, and totipotent differentiated cells: number of divisions is limited and specialized in most cases

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Label the following terms in order from smallest to largest -Chromosome -Gene -Genome -Adenine -Plasmid

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Smallest 1. Adenine 2. Gene 3. Plasmid 4. Chromosome 5. Genome Largest

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DNA Sequencing

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Determines the nucleotide sequence of DNA fragments.

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DNA probe

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A single-stranded sequence of DNA, labeled with radioactive isotope or fluorescent tag, used to detect the presence of a known sequence of nucleotides.

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Preimplantation Genetic Diagnosis

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Uses PCR and DNA probes to detect genetic diseases in fetuses, newborns, children, and adults.

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