3 therapeutic cloning

1. Types of Cloning 1 recombinant DNA technology or DNA cloning, the transfer of a DNA fragment of interest from one organism to a self-replicating genetic element such as a bacterial plasmid. The DNA of interest can then be propagated in a foreign host cell. 2 reproductive cloning Reproductive cloning is a technology used to generate an animal that has the same nuclear DNA as another currently or previously existing animal. In a process called "somatic cell nuclear transfer" (SCNT), genetic material from the nucleus of a donor adult cell is transferred to an egg whose nucleus, and thus its genetic material, has been removed. 3 therapeutic cloning "embryo cloning," is the production of human embryos for use in research. The goal of this process is not to create cloned human beings, but rather to harvest stem cells that can be used to study human development and to treat disease.

2. Applications 1 recombinant DNA technology or DNA cloning, This type of cloning is important for learning about other related technologies, such as gene therapy, genetic engineering of organisms, and sequencing genomes Gene therapy can be used to treat certain genetic conditions by introducing virus vectors that carry corrected copies of faulty genes into the cells of a host organism. Genes from different organisms that improve taste and nutritional value or provide resistance to particular types of disease can be used to genetically engineer food crops. 2 reproductive cloning Reproductive cloning also could be used to repopulate endangered animals or animals that are difficult to breed 3 therapeutic cloning Therapeutic cloning technology may some day be used in humans to produce whole organs from single cells or to produce healthy cells that can replace damaged cells in degenerative diseases such as Alzheimer's or Parkinson's.

3. Stem cells—Cells with the ability to divide for indefinite periods in culture and to give rise to specialized cells. Embryonic stem cells—Primitive (undifferentiated ) cells derived from a 5-day preimplantation embryo that are capable of dividing without differentiating for a prolonged period in culture, and are known to develop into cells and tissues of the three primary germ layers. Somatic (adult) stem cells—A relatively rare undifferentiated cell found in many organs and differentiated tissues with a limited capacity for both self renewal (in the laboratory) and differentiation. Such cells vary in their differentiation capacity, but it is usually limited to cell types in the organ of origin. This is an active area of investigation. Induced pluripotent stem cells—Somatic (adult) cells reprogrammed to enter an embryonic stem cell–like state by being forced to express factors important for maintaining the "stemness" of embryonic stem cells (ESCs). Scientists are actively comparing iPSCs and ESCs to identify important similarities and differences.

4. Adult Vs. Embryonic One major difference between adult and embryonic stem cells is their different abilities in the number and type of differentiated cell types they can become. Embryonic stem cells can become all cell types of the body because they are pluripotent. Adult stem cells are thought to be limited to differentiating into different cell types of their tissue of origin.

5. Stem cells are distinguished from other cell types by two important characteristics. First, they are unspecialized cells capable of renewing themselves through cell division, sometimes after long periods of inactivity. Second, under certain physiologic or experimental conditions, they can be induced to become tissue- or organ-specific cells with special functions. In some organs, such as the gut and bone marrow, stem cells regularly divide to repair and replace worn out or damaged tissues. In other organs, however, such as the pancreas and the heart, stem cells only divide under special conditions.

6. DNA Technologies Cloning, Sequencing and DNA Microarrays 1. Cloning Article Analysis TEXT= 366 to 368 2. Cloning techniques, recombinant DNA cloning animation TEXT= 373, 367and 368 3.DNA Microarrays TEXT= 375to 379 Review your cloning knowledge The Genetic Science Learning Center http://learn.genetics.utah.edu/content/tech/cloning/whatiscloning/ Read over the steps of somatic cell nuclear transfer (SCNT) and then view the two animations comparing Natural reproduction to SCNT. • Summarize the difference between Natural reproduction and SCNT • Describe what is done to the cell called the “ egg cell donor” in SCNT

7. Cloning Continued • Today you will complete the “Click and Clone” activity from the Utah Genetics web page • Answer the questions provided on the worksheet as you try your hand at cloning for yourself. Once you are done turn in the worksheet and review the information provided in your packet on “DNA sequencing”. Answer the following questions about this process What can the sequence of a gene be used to predict? In sequencing what will cause the synthesis of a new strand to stop? What is the result of this halt in strand synthesis? How do we determine the sequence of the original DNA template at the end of the reaction? Have the answers to these questions completed on a sheet of notebook paper for Monday

8. Sequencing Now that you have read a bit on the process of sequencing we will take a look at the steps of this procedure http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/D/DNAsequencing.html Review the information on this web page concerning the steps of sequncing . There are many steps so you may want to take some notes to help you keep track of this process. Q: What is the difference between normal (deoxy) nucleotides and dideoxynucleotides Q: What is the other name for the dideoxy method? Why? Once you have reviewed this web page and answered the questions above, click on “Animation of this proccedure”. Answer the questions provided on the worksheet as you go through the steps of DNA sequencing.

9. Sequencing continued

10. DNA Microarrays Now that you have worked with the process of making and analyzing an array, we will take a look at the social impact of such technologies. Read the articles found at the 3 web addresses listed below to learn about DNA Dragnets, how they are used and their impact on society. These articles contain some information that is purely opinion. This information should help you to form your own opinion on the use of this technology. 60 minutes article http://www.cbsnews.com/stories/2004/09/10/60minutes/main642684.shtml time article http://www.time.com/time/magazine/article/0,9171,1018083,00.html Christian science monitor article... against dragnets http://www.csmonitor.com/2003/0221/p03s01-usju.html Finally, read the printed out article provided to you concerning Dragnets. Fill out the social impact decision form. Once you have done this, write a response to what you have read today. Follow the guidelines provided on the next slide to help you.

11. Response I am not so much concerned with length… as a well thought out response to what you have read. • Requirements: • A 2 to 3 page response to what you have read concerning dragnet use. Include your personal opinion on the use of this technology and back this opinion up with referenced sources. • This is to be typed, double space, no more than 14 font and no print larger than Ariel please. • This should include a bibliography page. You may use any of the sources provided to you today and as many sources as you would like, provided you use a minimum of 3 sources. One source must be a source I did not provide you with (It can not be the article or 3 web sources you looked at today) • You are to use the guidelines set forth by the English department for the completion of this assignment. These can be found on the OCHS web page they are the links titled: Research Paper Style Guidelines and The Works Cited Guidelines Sheet. Please e-mail me if you have questions concerning either of these documents. • Please turn in a printed copy and an electronic copy of this assignment to me on Monday 3-1-10 in or before class. Do not depend on doing all of this in class… if something does not work correctly and you are unable to print it out or e-mail it to me… your assignment will be marked late.

12. Review Topics we have covered include: Cloning types and techniques Sequencing steps and results Microarray procedure and application DNA Dragnet use and social impact Plasmid Cloning Methods Investigation http://www.sumanasinc.com/webcontent/animations/content/plasmidcloning.html “Cycle Sequencing” on Dolan DNA Learning Center http://www.dnalc.org/resources/animations/cycseq.html http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/D/DNAsequencing.html Cloning Article responses http://www.ornl.gov/sci/techresources/Human_Genome/elsi/cloning.shtml http://stemcells.nih.gov/info/basics/basics1.asp

13. “Click and Clone” on Utah Genetics http://learn.genetics.utah.edu/content/tech/cloning/ “Sequence For Yourself” http://www.pbs.org/wgbh/nova/genome/sequencer.html#

14. How does SCNT differ from the natural way of making an embryo? The fertilization of an egg by a sperm and the SCNT cloning method both result in the same thing: a dividing ball of cells, called an embryo. So what exactly is the difference between these methods? An embryo is composed of cells that contain two complete sets of chromosomes. The difference between fertilization and SCNT lies in where those two sets originated. In fertilization, the sperm and egg both contain one set of chromosomes. When the sperm and egg join, the resulting zygote ends up with two sets - one from the father (sperm) and one from the mother (egg). In SCNT, the egg cell's single set of chromosomes is removed. It is replaced by the nucleus from a somatic cell, which already contains two complete sets of chromosomes. Therefore, in the resulting embryo, both sets of chromosomes come from the somatic cell.