Nuclear transfer

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BTX ECM 2001 Electrofusion generator used for nuclear transfer applications

Nuclear transfer is a form of cloning. The step involves removing the DNA from an oocyte (unfertilised egg), and injecting the nucleus which contains the DNA to be cloned. In rare instances, the newly constructed cell will divide normally, replicating the new DNA while remaining in a pluripotent state. If the cloned cells are placed in the uterus of a female mammal, a cloned organism develops to term in rare instances. This is how Dolly the Sheep and many other species were cloned. Cows are commonly cloned to select those that have the best milk production. On 24 January 2018, two monkey clones were reported to have been created with the technique for the first time. [1] [2] [3]

Contents

Despite this, the low efficiency of the technique has prompted some researchers, notably Ian Wilmut, creator of Dolly the cloned sheep, to abandon it. [4]

Tools and reagents

Nuclear transfer is a delicate process that is a major hurdle in the development of cloning technology. [5] Materials used in this procedure are a microscope, a holding pipette (small vacuum) to keep the oocyte in place, and a micropipette (hair-thin needle) capable of extracting the nucleus of a cell using a vacuum. For some species, such as mouse, a drill is used to pierce the outer layers of the oocyte.

Various chemical reagents are used to increase cloning efficiency. Microtubule inhibitors, such as nocodazole, are used to arrest the oocyte in M phase, during which its nuclear membrane is dissolved. Chemicals are also used to stimulate oocyte activation. When applied the membrane is completely dissolved.

Somatic cell nuclear transfer

Somatic Cell Nuclear Transfer (SCNT) is the process by which the nucleus of an oocyte (egg cell) is removed and is replaced with the nucleus of a somatic (body) cell (examples include skin, heart, or nerve cell). The two entities fuse to become one and factors in the oocyte cause the somatic nucleus to reprogram to a pluripotent state. The cell contains genetic information identical to the donated somatic cell. After stimulating this cell to begin dividing, in the proper conditions an embryo will develop. Stem cells can be extracted 5–6 days later and used for research. [6]

Reprogramming

Genomic reprogramming is the key biological process behind nuclear transfer. Currently unidentified reprogramming factors present in oocytes are capable of initiating a cascade of events that can reset the mature, specialized cell back to an undifferentiated, embryonic state. These factors are thought to be mainly proteins of the nucleus.

See also

Related Research Articles

<span class="mw-page-title-main">Cloning</span> Process of producing individual organisms with identical genomes

Cloning is the process of producing individual organisms with identical genomes, either by natural or artificial means. In nature, some organisms produce clones through asexual reproduction; this reproduction of an organism by itself without a mate is known as parthenogenesis. In the field of biotechnology, cloning is the process of creating cloned organisms of cells and of DNA fragments.

<span class="mw-page-title-main">Dolly (sheep)</span> First cloned mammal (1996–2003)

Dolly was a female Finn-Dorset sheep and the first mammal that was cloned from an adult somatic cell. She was cloned by associates of the Roslin Institute in Scotland, using the process of nuclear transfer from a cell taken from a mammary gland. Her cloning proved that a cloned organism could be produced from a mature cell from a specific body part. Contrary to popular belief, she was not the first animal to be cloned.

<span class="mw-page-title-main">Human cloning</span> Creation of a genetically identical copy of a human

Human cloning is the creation of a genetically identical copy of a human. The term is generally used to refer to artificial human cloning, which is the reproduction of human cells and tissue. It does not refer to the natural conception and delivery of identical twins. The possibilities of human cloning have raised controversies. These ethical concerns have prompted several nations to pass laws regarding human cloning.

<span class="mw-page-title-main">Stem cell</span> Undifferentiated biological cells that can differentiate into specialized cells

In multicellular organisms, stem cells are undifferentiated or partially differentiated cells that can differentiate into various types of cells and proliferate indefinitely to produce more of the same stem cell. They are the earliest type of cell in a cell lineage. They are found in both embryonic and adult organisms, but they have slightly different properties in each. They are usually distinguished from progenitor cells, which cannot divide indefinitely, and precursor or blast cells, which are usually committed to differentiating into one cell type.

<span class="mw-page-title-main">Somatic cell nuclear transfer</span> Method of creating a cloned embryo by replacing the egg nucleus with a body cell nucleus

In genetics and developmental biology, somatic cell nuclear transfer (SCNT) is a laboratory strategy for creating a viable embryo from a body cell and an egg cell. The technique consists of taking an denucleated oocyte and implanting a donor nucleus from a somatic (body) cell. It is used in both therapeutic and reproductive cloning. In 1996, Dolly the sheep became famous for being the first successful case of the reproductive cloning of a mammal. In January 2018, a team of scientists in Shanghai announced the successful cloning of two female crab-eating macaques from foetal nuclei.

<span class="mw-page-title-main">Germ cell</span> Gamete-producing cell

A germ cell is any cell that gives rise to the gametes of an organism that reproduces sexually. In many animals, the germ cells originate in the primitive streak and migrate via the gut of an embryo to the developing gonads. There, they undergo meiosis, followed by cellular differentiation into mature gametes, either eggs or sperm. Unlike animals, plants do not have germ cells designated in early development. Instead, germ cells can arise from somatic cells in the adult, such as the floral meristem of flowering plants.

<span class="mw-page-title-main">Embryonic stem cell</span> Type of pluripotent blastocystic stem cell

Embryonic stem cells (ESCs) are pluripotent stem cells derived from the inner cell mass of a blastocyst, an early-stage pre-implantation embryo. Human embryos reach the blastocyst stage 4–5 days post fertilization, at which time they consist of 50–150 cells. Isolating the inner cell mass (embryoblast) using immunosurgery results in destruction of the blastocyst, a process which raises ethical issues, including whether or not embryos at the pre-implantation stage have the same moral considerations as embryos in the post-implantation stage of development.

<span class="mw-page-title-main">John Gurdon</span> English developmental biologist (born 1933)

Sir John Bertrand Gurdon is a British developmental biologist, best known for his pioneering research in nuclear transplantation and cloning.

Sir Ian Wilmut was a British embryologist and the chair of the Scottish Centre for Regenerative Medicine at the University of Edinburgh. He is best known as the leader of the research group that in 1996 first cloned a mammal from an adult somatic cell, a Finnish Dorset lamb named Dolly.

Polly and Molly, two ewes, were the first mammals to have been successfully cloned from an adult somatic cell and to be transgenic animals at the same time. This is not to be confused with Dolly the Sheep, the first animal to be successfully cloned from an adult somatic cell where there wasn’t modification carried out on the adult donor nucleus. Polly and Molly, like Dolly the Sheep, were cloned at the Roslin Institute in Edinburgh, Scotland.

In biology, reprogramming refers to erasure and remodeling of epigenetic marks, such as DNA methylation, during mammalian development or in cell culture. Such control is also often associated with alternative covalent modifications of histones.

Megan and Morag, two domestic sheep, were the first mammals to have been successfully cloned from differentiated cells. They are not to be confused with Dolly the sheep which was the first animal to be successfully cloned from an adult somatic cell or Polly the sheep which was the first cloned and transgenic animal. Megan and Morag, like Dolly and Polly, were cloned at the Roslin Institute in Edinburgh, Scotland in 1995.

<span class="mw-page-title-main">Induced pluripotent stem cell</span> Pluripotent stem cell generated directly from a somatic cell

Induced pluripotent stem cells are a type of pluripotent stem cell that can be generated directly from a somatic cell. The iPSC technology was pioneered by Shinya Yamanaka and Kazutoshi Takahashi in Kyoto, Japan, who together showed in 2006 that the introduction of four specific genes, collectively known as Yamanaka factors, encoding transcription factors could convert somatic cells into pluripotent stem cells. Shinya Yamanaka was awarded the 2012 Nobel Prize along with Sir John Gurdon "for the discovery that mature cells can be reprogrammed to become pluripotent."

<span class="mw-page-title-main">Shinya Yamanaka</span> Japanese stem cell researcher

Shinya Yamanaka is a Japanese stem cell researcher and a Nobel Prize laureate. He is a professor and the director emeritus of Center for iPS Cell Research and Application, Kyoto University; as a senior investigator at the UCSF-affiliated Gladstone Institutes in San Francisco, California; and as a professor of anatomy at University of California, San Francisco (UCSF). Yamanaka is also a past president of the International Society for Stem Cell Research (ISSCR).

Keith Henry Stockman Campbell was a British biologist who was a member of the team at Roslin Institute that in 1996 first cloned a mammal, a Finnish Dorset lamb named Dolly, from fully differentiated adult mammary cells. He was Professor of Animal Development at the University of Nottingham. In 2008, he received the Shaw Prize for Medicine and Life Sciences jointly with Ian Wilmut and Shinya Yamanaka for "their works on the cell differentiation in mammals".

<span class="mw-page-title-main">Cell potency</span> Ability of a cell to differentiate into other cell types

Cell potency is a cell's ability to differentiate into other cell types. The more cell types a cell can differentiate into, the greater its potency. Potency is also described as the gene activation potential within a cell, which like a continuum, begins with totipotency to designate a cell with the most differentiation potential, pluripotency, multipotency, oligopotency, and finally unipotency.

<span class="mw-page-title-main">Partial cloning</span> Method in cell biology

In the field of cell biology, the method of partial cloning (PCL) converts a fully differentiated old somatic cell into a partially reprogrammedyoung cell that retains all the specialised functions of the differentiated old cell but is simply younger. The method of PCL reverses characteristics associated with old cells. For example, old, senescent, cells rejuvenated by PCL are free of highly condensed senescence-associated heterochromatin foci (SAHF) and re-acquire the proliferation potential of young cells. The method of PCL thus rejuvenates old cells without de-differentiation and passage through an embryonic, pluripotent, stage.

Induced stem cells (iSC) are stem cells derived from somatic, reproductive, pluripotent or other cell types by deliberate epigenetic reprogramming. They are classified as either totipotent (iTC), pluripotent (iPSC) or progenitor or unipotent – (iUSC) according to their developmental potential and degree of dedifferentiation. Progenitors are obtained by so-called direct reprogramming or directed differentiation and are also called induced somatic stem cells.

<span class="mw-page-title-main">Zhong Zhong and Hua Hua</span> Worlds first cloned primates (born 2017)

Zhong Zhong and Hua Hua are a pair of identical crab-eating macaques that were created through somatic cell nuclear transfer (SCNT), the same cloning technique that produced Dolly the sheep in 1996. They are the first cloned primates produced by this technique. Unlike previous attempts to clone monkeys, the donated nuclei came from fetal cells, not embryonic cells. The primates were born from two independent surrogate pregnancies at the Institute of Neuroscience of the Chinese Academy of Sciences in Shanghai.

Transflammation describes the process by which innate immune response mechanisms affect the epigenetic plasticity of a cell during nuclear reprogramming. This phenomenon is essential in dedifferentiating a somatic cell to a pluripotent cell and also in transdifferentiating a terminally differentiated cell to another terminally differentiated cell.

References

  1. Liu, Zhen; et al. (24 January 2018). "Cloning of Macaque Monkeys by Somatic Cell Nuclear Transfer". Cell . 172 (4): 881–887.e7. doi: 10.1016/j.cell.2018.01.020 . PMID   29395327.
  2. Briggs, Helen (24 January 2018). "First monkey clones created in Chinese laboratory". BBC News . Retrieved 24 January 2018.
  3. "Scientists Successfully Clone Monkeys; Are Humans Up Next?". The New York Times . Associated Press. 24 January 2018. Retrieved 24 January 2018.
  4. Roger Highfield Dolly creator Prof Ian Wilmut shuns cloning. The Telegraph. 16 November 2007
  5. "PNAS Classics -- Nuclear Transfer". www.pnas.org. Archived from the original on 2003-11-21.
  6. Tachibana, M.; Amato, P.; Sparman, M.; Gutierrez, N. M.; Tippner-Hedges, R.; Ma, H.; Kang, E.; Fulati, A.; Lee, H. S.; Sritanaudomchai, H.; Masterson, K.; Larson, J.; Eaton, D.; Sadler-Fredd, K.; Battaglia, D.; Lee, D.; Wu, D.; Jensen, J.; Patton, P.; Gokhale, S.; Stouffer, R. L.; Wolf, D.; Mitalipov, S. (2013). "Human Embryonic Stem Cells Derived by Somatic Cell Nuclear Transfer". Cell. 153 (6): 1228–38. doi:10.1016/j.cell.2013.05.006. PMC   3772789 . PMID   23683578.