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

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Biology

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Links Among Engineering, Technology, Science, and Society

BIO1.ETS2.3

Reproductive Cloning

Klonowanie reprodukcyjne to produkcja genetycznie identycznej kopii, klonu organizmu. Owca Dolly, wyprodukowana w 1996 roku, była pierwszym klonem dorosłego zwierzęcia, ale od tego czasu sklonowano kilka innych gatunków, w tym psy i koty.
Najczęstszą metodą klonowania dorosłych zwierząt jest transfer jądra komórki somatycznej. Najpierw jądro jest usuwane z komórki jajowej, a następnie od zwierzęcia, które ma zostać sklonowane, pobiera się komórkę somatyczną, taką jak komórka skóry. Jądro zawierające chromosomalne DNA jest usuwane i wstrzykiwane do komórki jajowej. Komórka jajowa jest stymulowana do podziału przez obróbkę chemiczną lub elektryczną i tworzy zarodek, który jest zagnieżdżany w macicy dorosłej samicy, gdzie rozwija się aż do urodzenia.
Powstały klon ma identyczne chromosomalne DNA jak oryginalne zwierzę, ale mitochondrialne DNA jest często inne, ponieważ mitochondria pochodzą z cytoplazmy komórki jajowej, zwykle od innego osobnika.
Również różnice fenotypowe między klonem a oryginałem mogą wystąpić z powodu czynników środowiskowych i epigenetycznych podczas rozwoju. Tak jak bliźnięta jednojajowe nieco różnią się od siebie, mimo że mają to samo DNA.

Ayala, Francisco J. "Klonowanie ludzi? Rozważania biologiczne, etyczne i społeczne." Proceedings of the National Academy of Sciences of the United States of America 112, nr 29 (21 lipca 2015): 8879–86. [<a href="https://doi.org/10.1073/pnas.1501798112" target="__blank">Źródło</a>]
Keefer, Carol L. "Sztuczne klonowanie zwierząt domowych". Proceedings of the National Academy of Sciences of the United States of America 112, nr 29 (21 lipca 2015): 8874–78. <a href="https://doi.org/10.1073/pnas.1501718112" target="__blank">https://doi.org/10.1073/pnas.1501718112</a>.

klonowanie reprodukcyjne

Klonowanie reprodukcyjne to proces wytwarzania genetycznie identycznej kopii — klonu — całego organizmu. Chociaż klony można wytwarzać poprzez podział wczesnego zarodka—w sposób podobny do tego, co dzieje się naturalnie w przypadku bliźniąt jednojajowych — klonowanie dorosłych zwierząt zwykle odbywa się w procesie zwanym transferem jądra komórki somatycznej (SCNT).

<h4>Transfer jądra komórek somatycznych</h4>

W SCNT od zwierzęcia pobiera się komórkę jajową i usuwa się jej jądro, tworząc jajo pozbawione jądra. Następnie od zwierzęcia przeznaczonego do klonowania pobiera się komórkę somatyczną — każdą komórkę niebędącą komórką płciową—Jądro komórki somatycznej jest następnie przenoszone do pozbawionego jądra jaja—albo przez bezpośrednie wstrzyknięcie, albo przez fuzję komórki somatycznej z komórką jajową za pomocą prądu elektrycznego.

Jajo zawiera teraz jądro wraz z chromosomalnym DNA zwierzęcia, które ma zostać sklonowane. Pobudza się go do podziału, tworząc zarodek, który następnie wszczepia się do macicy matki zastępczej. Jeśli wszystko pójdzie dobrze, rozwija się normalnie i rodzi się klon.

Chociaż proces ten stosowano do skutecznego klonowania wielu różnych typów zwierząt—w tym owiec, krów, mułów, królików i psów—jego wskaźnik powodzenia jest niski i tylko niewielki procent zarodków przeżywa do porodu. Sklonowane zwierzęta, które przeżyją do urodzenia, również wydają się starzeć i przedwcześnie umierać. Dzieje się tak, ponieważ ich DNA pochodzi z dorosłych komórek, które uległy skróceniu telomerów — utracie niewielkiej części ochronnych końców chromosomów przy każdym podziale komórki — w ramach normalnego procesu starzenia.

Chociaż chromosomalny DNA klonu jest taki sam jak DNA dawcy jądra, może mieć inny mitochondrialny DNA, ponieważ mitochondria pochodzą z cytoplazmy komórki jajowej, która zwykle pochodzi od innego zwierzęcia. Mogą również wystąpić różnice fenotypowe między klonem a pierwotnym zwierzęciem, ze względu na czynniki środowiskowe i epigenetyczne. Na przykład pierwszy sklonowany kot, Cc, wyglądał zupełnie inaczej niż pierwotny kot, ponieważ wzór sierści wynika z losowej inaktywacji chromosomu X w różnych komórkach.

Pomimo wyzwań technicznych klonowanie reprodukcyjne ma wiele potencjalnych zastosowań, w tym produkcję genetycznie identycznych zwierząt badawczych, zwierząt gospodarskich o pożądanych cechach i potomstwa gatunków zagrożonych. Ma nawet potencjalne zastosowanie w leczeniu niepłodności i chorób u ludzi, chociaż nie przeprowadzono jeszcze klonowania ludzi, co budziłoby wątpliwości etyczne.

Klonowanie reprodukcyjne

Klonowanie reprodukcyjne to proces wytwarzania genetycznie identycznej kopii – klonu – całego organizmu. Podczas gdy klony mogą być produkowane przez ...

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Kluczowe terminy i pojęcia

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...

crispr

CRISPR-Cas9 Gene Editing in Action

CRISPR-Cas9 is a genome editing tool that lets scientists change DNA at a specific location. It can add, remove, or rearrange genetic material. Researchers study it because it may help treat genetic disorders such as hemophilia and sickle cell anemia. CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats, and Cas9 is a CRISPR-associated protein 9. The basic system includes a Cas9 endonuclease, which is an enzyme that cuts DNA, and a small RNA that guides Cas9 to the...

gene therapy

Gene Delivery for Serious Diseases

Gene therapy is a technique used to treat serious disease by adding a gene to a person’s cells. The added gene may be a healthy copy of a mutated gene. It may also be a different gene that inactivates or compensates for the disease-causing gene. One example is severe combined immunodeficiency, or SCID, caused by a mutation in the gene for adenosine deaminase. In that case, a working copy of the gene can be inserted so the patient’s cells make the enzyme again. This can cure the disease in some...

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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Obtain, evaluate, and communicate information on how molecular biotechnology may be used in a variety of fields.
EXPLANATION: The goal is to help students appreciate that the basic science knowledge being studied can be manipulated and used to design useful tools for further scientific investigations, medical treatments, agricultural yields, etc. A detailed understanding of any technical procedures is not expected, but rather a "big picture" view of concepts the technology utilizes and the applications the technology is being used for. These technologies may be best investigated along with the standards from LS1 From Molecules to Organisms: Structure and Functions, as the students are learning about DNA and protein. Comparative DNA and protein sequence analysis is also a large part of current phylogenic research and could accompany LS4 Biological Change: Unity and Diversity. Students may also consider the pros and cons of biotechnical applications. Investigation of techniques utilized could come from field trips to witness the use of these techniques in practice, lab exercises, virtual labs, simulation activities, interviews with professionals, etc. Molecular techniques could include: PCR, electrophoresis, restriction enzyme digestion of DNA, DNA sequencing, plasmid-based transformation, transfection, etc. These techniques are used in fields of medicine, agriculture, biomedical engineering, forensic science, etc.

BIO1.ETS2.1

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Investigate means by which karyotypes are utilized in diagnostic medicine.
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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