Altered Nuclear Transfer ANT

Maria Ssozi 3-2-13

The ANT procedure has been proposed as morally acceptable and is also safe way to obtain stem cells without the creation and destruction of human embryos. ANT was originally proposed by Dr. William B. Hurlbut, a Stanford University professor and a member of the President Council on bioethics. At Whitehead Institute for Biomedical Research scientist have demonstrated a theoretical--and controversial technique for generating embryonic stem cells. Most of the scientists first tests were on mice, and first successful with mice. Altered Nuclear Transfer (ANT) advices scientists should first create genetically altered embryos that are not able to implant in a uterus and take stem cells from these embryos, since they have no human potential. Altered nuclear transfer is when an unfertilized egg is retrieved by the penetrating of the outside layer using a small needle. After the cell is penetrated a microscopic needle retrieves the nucleus, which contains the most genetic information, and from the donor egg. The microscopic needle then places sperm and eggs from the intended parents into the egg, an impulse charged is then used instead of normal fertilization, and the embryo grows and divides. ANT is also a dignified alternative to somatic cell nuclear transfer (SCNT), which mostly known as therapeutic cloning. In SCNT, a donor nucleus taken from, for example a skin cell, and placed into a donor egg cell where the nucleus was just removed. The egg is manipulated into thinking it has been fertilized, this causes it to grow into a blastocyst, the early stages of the mammalian embryo, containing an inner cell mass, a cavity and an outer layer, with a mass of about 100 cells. This is where the stem cells are removed, these stem cells have the ability to divide and replicate themselves indefinitely, and the cells can also form any type of tissue in the body. The only way to gather/extract these cells is to destroy the blastocyst. The difference between ANT and SCNT is that in ANT before the donor nucleus is switched into the egg cell, its DNA is changed so the resulting blastocyst can never be an embryo. Through this process the potential human being will not be destroyed once the cells are extracted. Scientists disable the Cdx2 in mouse cells in order to create a blastocyst that cant implant in a uterus. Cdx2 is a gene thats member of the caudal-related homeobox transcription factor gene family, it is also an encoded protein which regulates intestine-specific genes that are involved with cell growth and differentiation. Cdx2 plays a role in the beginning stages of embryonic development of the intestinal tract as well. An abnormal expression of this gene is connected with intestinal inflammation and tumor genesis. Scientists accomplished creating a blastocyst that cant reproduce by using a technique called RNA interference (RNAi), a biological process where RNA molecules inhibit gene expression, by causing the destruction of specific mRNA molecules. In RNAi short interfering RNA (siRNA) molecules are shaped to target an individual gene and agitate its competence to make protein, because of this the gene is shut off. Scientists Jaenisch and Meissner designed a particular form of siRNA that shuts off the gene in the donor nucleus and incorporates itself into all cells in the blastocyst; as a result all of the mouse blastocysts were incapable of implanting themselves.

Although the stem cells were extracted from the blastocysts, but the Cdx2 was still disabled in each of the new cells. To fix this problem scientist, Meissner deleted siRNA molecule by changing a plasmid, unit of DNA that can replicate in a cell apart from the nucleus, into each cell. The cells came from this experiment proved to be just as strong and flexible as the stem cells that were taken the traditional way. This analysis not only explains what is morally troubling about CBR (chemical, bacteriological, or radiological) but it also suggests a resolution to our national policy impasse over its use. What CBR produces also fails to harvest ESCs (embryonic stem cells) in a fully constituted, a human embryo. But by altering the CBR process could be able to produce ESCs from a biologic construct that only have partial developmental potential, so that no embryo would be created or destroyed. The analysis of this approach was to lead a technological solution for the conflict over Embryonic Stem Cell research. In regular nuclear transfer, the cell nucleus is removed from a somatic cell and transfers it into an oocyte that has had its own nucleus removed. The oocyte then gets a full complement of DNA and after it is electrically stimulated, starts to divide like a naturally fertilized egg. ANT uses the technology of nuclear transfer except for using a pre-emptive alteration that assures that no embryo is created. The somatic cell nucleus or the enucleated oocyte's contents are first changed before the somatic cell nucleus is transferred into the egg. The alterations cause the somatic cell DNA to function in such a way that no embryo is created, but pluripotent stem cells are made. In a January 2006 Jaenisch established the scientific feasibility of one example of the ANT approach in a series of experiments in which he obtained fully functional ESCs from a construct that is radically different in developmental potential than a human embryo. ANT-OAR, another variation of ANT standing for oocyte assisted reprogramming (ANT-OAR) has been used by Markus Grompe, Director of the Stem Cell Center at Oregon Health Science University. In this variation of ANT, changes of the nucleus of the adult body cell and the enucleated egg's contents before nuclear transfer would be forced into early expression of genes characteristic of a later and more specialized cell type. This cell type would be capable of producing pluripotent stem cells. The advantages of ANT are providing a uniquely flexible tool for embryonic stem cell research. Embryos left over from IVF procedures represent a limited pool of genotypes, the genomes of these embryos have never proven their capacity to form an organism because of mutations, recombinations and re-assortment of alleles in game to genesis. Embryonic stem cells produced by ANT, on the other hand, would have genotypes of proven potential. ANT could give a full range of genotypes, of which include specific genetic types for tissuecompatible transplantation. This technique would also offer a wider range of scientific and medical possibilities than ESC lines made from left over IVF embryos, including generation of diverse and pre-designed ESC lineages for disease modeling and pharmaceutical development. ANT would also unburden ESC research from the additional ethical concerns of the left over' IVF embryos, of which include the attendance of clinical and legal complexities in a level great personal and social sensitivity. The last link with IVF, the procurement of ocytes, is a subject of intense scientific research and appears to have several prospects for obtaining eggs without the morally dubious and expensive super-ovulation of female patients. The proposal for altered nuclear transfer, developed by a wide consultation of leading scientists, has moral

philosophers, and religious authorities, representing a third option technological solution to the current moral impasse concerning the destruction of human embryos to obtain embryonic stem cells. Using the techniques of somatic-cell nuclear transfer scientists could construct a biologic entity from its very inception, lacking the attributes and capacities of a human embryo. Experiments with mice already show evidence that we may be able to generate functional embryonic stem cells from a system that is not an organism but is biologically more akin to tissue or cell culture. There is a natural precedent for entities that lack the characteristics of organisms, but can generate embryonic stem cells or a functional equivalent. Teratomas are germ-cell tumors that generate all three primary embryonic germ-layer cell types, they also have more advanced cells and tissues. These chaotic, disorganized, and nonfunctional masses entirely lack the structural and dynamic character of an organism.

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