Name: cloning animals with scnt
Uploaded: 2019-03-11T09:21:22.000Z
Duration: 1 min 27 s
Channel: JoVE

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Links Among Engineering, Technology, Science, and Society

BIO1.ETS2.3

Reproductive Cloning

- [Anlatıcı] Üreme amaçlı klonlama, bir organizmanın genetik olarak kopyasını, klonunu üretme işidir.
 1996'da üretilen koyun Dolly, yetişkin bir hayvanın ilk klonuydu.
 Ama o zamandan bu yana başka türler de klonlandı, buna kedi ve köpekler de dâhil.
Yetişkin hayvanları klonlamanın en yaygın yöntemi, somatik hücre çekirdeği transferidir.
 Önce çekirdek yumurta hücresinden çıkarılır.
 Sonra deri hücresi gibi bir tür somatik hücre, klonlanacak hayvandan alınır.
Kromozomal DNA'yı taşıyan çekirdek çıkarılır ve yumurtaya enjekte edilir.
 Yumurta, kimyasal ya da elektriksel müdahaleyle bölünmeye teşvik edilir ve embriyo oluşturur.
 Embriyo yetişkin dişinin rahmine yerleştirilir ve orada doğuma kadar gelişmeyi sürdürür.
Sonuçta ortaya çıkan klonun kromozomal DNA'sı orijinal hayvanla aynıdır ancak mitokondriyal DNA'sı genelde farklıdır.
 Çünkü mitokondri yumurtanın sitoplazmasından, genelde başka bir bireyden gelir.
 Ayrıca klon ve orijinal arasında gelişim sırasındaki çevresel ve epigenetik faktörler sebebiyle fenotipik farklılıklar da olabilir.
Bu, tek yumurta ikizlerinin aynı DNA'yı taşımalarına rağmen biraz farklı olmalarına benzer.

Ayala, Francisco J. "İnsanları Klonlamak? Biyolojik, Etik ve Sosyal Hususlar." Amerika Birleşik Devletleri Ulusal Bilimler Akademisi Bildirileri 112, no. 29 (21 Temmuz 2015): 8879–86. [<a href="https://doi.org/10.1073/pnas.1501798112" target="__blank">Kaynak</a>]
Keefer, Carol L. "Evcil Hayvanların Yapay Klonlanması." Amerika Birleşik Devletleri Ulusal Bilimler Akademisi Bildirileri 112, no. 29 (21 Temmuz 2015): 8874–78. <a href="https://doi.org/10.1073/pnas.1501718112" target="__blank">https://doi.org/10.1073/pnas.1501718112</a>.

Üreme klonlaması

Reprodüktif klonlama, tüm bir organizmanın genetik olarak kopyasını (bir klon) üretme işlemidir. Klonlar erken bir embriyonun bölünmesiyle üretilebilirken (tek yumurta ikizlerinde doğal olarak görülene benzer) yetişkin hayvanların klonlaması genellikle somatik hücre nükleer transferi (SHÇT) adı verilen bir işlemle yapılır.

<h4>Somatik Hücre Çekirdeği Transferi</h4>

 SHÇT'de, bir hayvandan bir yumurta hücresi alınır ve çekirdeği çıkarılarak çekirdeksiz bir yumurta oluşturulur. Daha sonra klonlanacak hayvandan somatik bir hücre (seks hücresi olmayan herhangi bir hücre) alınır. Somatik hücrenin çekirdeği daha sonra çekirdeksiz yumurtaya aktarılır; ya doğrudan enjeksiyonla ya da elektrik akımı kullanarak somatik hücrenin yumurtaya füzyonu ile.

 Yumurta artık klonlanacak hayvanın kromozomal DNA'sına sahip çekirdeğini içerir. Yumurta bölünmeye ve daha sonra taşıyıcı annenin rahmine yerleştirilen bir embriyo oluşturmaya teşvik edilir. Her şey yolunda giderse, normal şekilde gelişir ve klon doğar.

Bu işlem birçok farklı hayvan türünü (koyunlar, inekler, katırlar, tavşanlar ve köpekler dahil) başarılı bir şekilde klonlamak için kullanılmış olsa da, başarı oranı düşüktür ve embriyoların sadece küçük bir yüzdesi doğuma kadar hayatta kalmaktadır. Doğuma kadar hayatta kalan klonlanmış hayvanlar da erken dönemde yaşlanıp ölüyor gibi görünmektedir. Bunun nedeni, DNA'larının alındıkları canlıda normal yaşlanma sürecinin bir parçası olan telomer kısalması (her hücre bölünmesinde kromozomların koruyucu uçlarının küçük bir kısmının kaybı) sürecinden geçmiş olmasıdır.

Klonun kromozomal DNA'sı çekirdek donörünkiyle aynı olsa da, mitokondriler genellikle farklı bir hayvandan gelen yumurta hücresinin sitoplazmından geldiği için farklı mitokondriyal DNA'ya sahip olabilir. Ayrıca, çevresel ve epigenetik faktörler nedeniyle klon ve orijinal hayvan arasında fenotipik farklılıklar meydana gelebilir. Örneğin, ilk klonlanmış kedi Cc, orijinal kediden çok farklı görünüyordu, çünkü deri deseni farklı hücrelerdeki rastgele X kromozomu inaktivasyonundan kaynaklanıyordu.

Teknik zorluklara rağmen, reprodüktif klonlama genetik olarak aynı araştırma hayvanlarının üretimi, istenen özelliklere sahip hayvancılık ve nesli tükenmekte olan türlerin korunması da dahil olmak üzere birçok potansiyel kullanıma sahiptir. Hatta insanların klonlanması etik kaygılar sebebiyle henüz yapılmamış olmasına rağmen, infertilite ve farklı hastalıklarda potansiyel uygulamalara sahipti.

Üreme Klonlaması

Üreme klonlaması, tüm bir organizmanın genetik olarak aynı bir kopyasını (bir klonu) üretme sürecidir. Klonlar şu şekilde üretilebilirken...

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cloning animals with scnt

Reproductive cloning can produce a genetically identical copy of an entire organism. In adult animals, this is usually done with somatic cell nuclear transfer, or SCNT. Another way clones can form is by splitting an early embryo, which is similar to how identical twins arise naturally.

In SCNT, an egg cell is taken from an animal and its nucleus is removed. This creates an enucleated egg. A somatic cell, which is any body cell that is not a sex cell, is then taken from the animal to be cloned. The nucleus from that cell is placed into the enucleated egg by direct injection or by fusing the cells with an electrical current.

The egg now contains the chromosomal DNA of the animal being cloned. It is stimulated to divide and form an embryo. That embryo is then implanted into the uterus of a surrogate mother. If development proceeds normally, the clone is born.

This method has been used to clone many animals, including sheep, cows, mules, rabbits, and dogs. Even so, the success rate is low, and only a small percentage of embryos survive to birth. Clones that do survive may age and die early because the DNA comes from adult cells that already show telomere shortening, which is the loss of protective chromosome ends during cell division.

A clone has the same chromosomal DNA as the nucleus donor, but it may have different mitochondrial DNA. That is because mitochondria come from the egg cell cytoplasm, which usually comes from a different animal. Clones can also look different from the original animal because of environmental and epigenetic effects. The first cloned cat, Cc, is one example, since coat pattern depends on random X-chromosome inactivation in different cells.

Despite these challenges, reproductive cloning may be useful for making genetically identical research animals, livestock with desired traits, and offspring of endangered species. It may also have possible uses in human infertility and disease. However, humans have not been cloned, and doing so would raise ethical concerns.

Cloning Animals with SCNT

what is genetic engineering?

How Scientists Edit DNA for New Traits

Genetic engineering changes an organism’s DNA to add useful traits. Scientists use it in bacteria, plants, animals, and even viruses for research, medicine, agriculture, and industry. It can bring major benefits, but it also raises ethical concerns about humans and the food supply. This work is possible because DNA has the same basic structure and genetic code across life. That shared code lets scientists modify DNA in several ways. One method is CRISPR/Cas9, often called molecular scissors,...

dna isolation

Steps in DNA Extraction

DNA extraction is the process of removing and purifying DNA from cells. It is used in many biotechnology and research applications, including molecular cloning. The main goal is to free DNA from the cell and prepare it for later study or use. The first step is cell lysis, which means breaking cells open. Researchers may use a homogenizer, sonicator, or bead beater to physically lyse the cells. They also often add detergent to chemically disrupt the lipid-based cell membranes. This helps...

recombinant dna technology

Recombinant DNA in Cloning and GMOs

Recombinant DNA is made by joining DNA from different sources in the lab. Scientists often use it to clone genes and study them in cells that are easy to handle, such as bacteria. Organisms that carry recombinant DNA are called genetically modified organisms, or GMOs. This technology gives scientists new tools for research, medicine, and agriculture. To build recombinant DNA, researchers place a gene of interest into a vector, which is a DNA carrier that moves foreign DNA into a host cell. The...

PCR Cycle for DNA Amplification

PCR is a laboratory method for making many copies of a DNA segment. It can turn a small amount of DNA into millions or even billions of copies in just a few hours. The process uses a heat-resistant DNA polymerase inside a machine called a thermocycler. PCR was developed by Kary Mullis in 1983, and he received the 1993 Nobel Prize in Chemistry for it. The method is fast, relatively inexpensive, and precise. It has become useful for molecular cloning, gene mutagenesis, pathogen detection, gene...

complementary dna

From mRNA to cDNA for Gene Studies

Complementary DNA, or cDNA, is made from messenger RNA (mRNA) so scientists can study which genes are active in cells and tissues. Only genes that are transcribed into mRNA are expressed. By extracting mRNA from cells, researchers can compare gene expression across samples. cDNA is produced by reverse transcription. This process copies mRNA into DNA, which is useful because mRNA does not contain introns, or non-coding regions, and other regulatory sequences. As a result, cDNA can help...

antibiotic selection

Antibiotic Resistance in Cloning

Antibiotic resistance genes help scientists find bacteria that took up a plasmid with a gene of interest. In genetic engineering, this resistance is used as a marker, not just as a medical concern. The same process also connects to the growing problem of resistant bacteria in the real world. Antibiotics, such as penicillin, stop bacterial growth or kill bacteria. When bacteria naturally have, or are engineered to carry, an antibiotic resistance gene, the antibiotic no longer works on them.

transgenic organisms

Creating Transgenic Plants and Animals

Transgenic organisms are genetically engineered to carry a transgene, which is a gene from a different species, in their genome. The inserted gene may be a new gene for that organism or a different version of one it already has. Scientists usually make transgenes with recombinant DNA and DNA cloning methods. Researchers use transgenic bacteria, plants, and animals to answer biological questions and to build useful tools. To make a transgenic organism, they first choose a transgenesis method,...

in-vitro mutagenesis (gene knockouts)

Knockout Mice and Gene Replacement

Knockout mice help researchers study gene function by showing what happens when a gene is turned off. These engineered animals are also useful for modeling human diseases such as cancer, Parkinson’s disease, and diabetes. A specific gene can be targeted by using a targeting vector, which is a piece of engineered DNA that replaces the normal gene and inactivates it. The vector has matching DNA sequences on both ends. These homologous sequences let the vector swap in for the target gene through...

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CRISPR-Cas9 Gene Editing in Action

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gene therapy

Gene Delivery for Serious Diseases

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embryonic stem cells

How Embryonic Stem Cells Become Specialized

Embryonic stem cells are pluripotent cells that have not yet specialized. Pluripotent means they can give rise to many different cell types in the body. Because of this, they are useful in science and medicine for making cells needed in research or for replacing cells lost to damage or disease. These cells come from the inner cell mass of a blastocyst, which is an early embryo about 3 to 5 days after fertilization in humans. At this stage, the embryo has not yet implanted in the uterus. Human...

adult stem cells

Adult Stem Cells in the Small Intestine

Adult stem cells help maintain tissues by replacing old or damaged cells. These undifferentiated cells divide to make more stem cells or progenitor cells, which then become mature, specialized cells. Small groups of adult stem cells are found in many tissues, including the bone marrow, brain, skin, and gut. The small intestine shows this process clearly. Its lining, called the epithelium, is renewed continuously by adult stem cells. It is also the fastest replaced tissue in the human body,...

induced pluripotent stem cells

Reprogramming Mature Cells into Stem Cells

Reprogramming mature cells into stem cells lets scientists turn specialized cells back into an unspecialized state. These cells are called induced pluripotent stem cells, or iPSCs. Like embryonic stem cells, iPSCs are pluripotent, which means they can form all cell types. To make iPSCs, researchers start with mature cells such as skin fibroblasts or blood cells. The cells are grown in culture, and genes for several transcription factors are delivered with a viral vector. The cells then make...

reproductive cloning

Reproductive cloning can produce a genetically identical copy of an entire organism. In adult animals, this is usually done with somatic cell nuclear transfer, or SCNT. Another way clones can form is by splitting an early embryo, which is similar to how identical twins arise naturally. In SCNT, an egg cell is taken from an animal and its nucleus is removed. This creates an enucleated egg. A somatic cell, which is any body cell that is not a sex cell, is then taken from the animal to be cloned.

genomics

Genomics and DNA Sequencing

Genomics studies the full set of genetic material in an organism. In humans, that genome includes the DNA carried in 23 pairs of chromosomes in the nucleus, along with mitochondrial DNA. Genomics looks at both coding DNA and non-coding DNA, which helps scientists study how living things are built and how they differ. This field is useful in many areas of biology and medicine. Scientists use genomics to build phylogenetic trees, improve crop productivity and sustainability, help with criminal...

key terms and concepts

Explore how somatic cell nuclear transfer creates cloned animals, why success is limited, and how cloning may support research, agriculture, and conservation.

Cloning Animals with SCNT | JoVE K12

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In molecular biology, ligation refers to the joining of two DNA fragments through the formation of a phosphodiester bond.  An enzyme known as a ligase ...

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Obtain, evaluate, and communicate information on how molecular biotechnology may be used in a variety of fields.
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BIO1.ETS2.1

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EXPLANATION: Engineers incorporate growing scientific knowledge in order to increase benefits to humans and decrease cost and risks. Karyotyping technology and its use in medical diagnosis can complement the standards from LS3: Hereditary: Inheritance, and Variation of Traits, as the students are learning about chromosomal organization of genomic information that determines one's phenotype. Analyze a large set of karyotypes to identify common patterns. Identify variations from the most commonly observed patterns and correlate them with patient phenotypes to propose a cause and effect relationship, discussing limitations of correlative data interpretation.

BIO1.ETS2.2

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EXPLANATION: The utilization of new technologies in any field of science is dependent on both economic and social factors. In addition to evaluating these factors, scientists must also consider long-term consequences that may not be initially apparent. The emphasis should be on the construction of a rationale argument that supports a position on the use of an application with ethical and social impact. Students should begin to appreciate the differences in ethical values that exist, and recognize that discussion of these values is imperative as knowledge and technology continue to advance, even when resolutions of differences can be rare. After investigation of a specific biotechnology application, students can write a position paper and/or participate in a classroom debate.

Cloning Animals with SCNT

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