Name: gene delivery for serious diseases
Uploaded: 2026-08-26T08:21:16.396Z
Duration: 1 min 22 s
Channel: JoVE

K12 Standards

Tennessee Academic Standards

High School

Biology

links-among-engineering-technology-science-and-society

Links Among Engineering, Technology, Science, and Society

BIO1.ETS2.3

Gene Therapy

A terapia g&#xE9;nica &#xE9; a inser&#xE7;&#xE3;o de um gene em c&#xE9;lulas individuais para prevenir ou tratar doen&#xE7;as graves. O gene inserido pode ser uma c&#xF3;pia saud&#xE1;vel de um gene mutado ou pode ser um gene diferente que neutraliza os efeitos de um gene defeituoso. Os genes podem ser entregues in vivo, diretamente em uma pessoa, ou ex vivo, em c&#xE9;lulas removidas do corpo, que s&#xE3;o ent&#xE3;o transplantados de volta.O novo gene &#xE9; inserido usando um vetor, normalmente um v&#xED;rus modificado que pode entrar em uma c&#xE9;lula e entregar o DNA ao n&#xFA;cleo. Alguns vetores integram o DNA no genoma enquanto outros permanecem fora do cromossomo. Os vetores cont&#xEA;m uma regi&#xE3;o promotora, para que o gene possa ser transcrito.O gene introduzido &#xE9; ent&#xE3;o traduzido em prote&#xED;na, fornecendo um tratamento para a doen&#xE7;a.

Dunbar, Cynthia E., Katherine A. High, J. Keith Joung, Donald B. Kohn, Keiya Ozawa e Michel Sadelain. "A terapia genética atinge a maioridade." Ciência 359, nº 6372 (12 de janeiro de 2018): eaan4672. [<a href="https://doi.org/10.1126/science.aan4672" target="__blank">Fonte</a>]

Anguela, Xavier M. e Katherine A. High. "Entrando na Era Moderna da Terapia Genética." Revisão Anual de Medicina 70, nº 1 (2019): 273–88. [<a href="https://doi.org/10.1146/annurev-med-012017-043332" target="__blank">Fonte</a>]

terapia genética

A terapia gen&#xE9;tica &#xE9; uma t&#xE9;cnica onde um gene &#xE9; inserido nas c&#xE9;lulas de uma pessoa para prevenir ou tratar uma doen&#xE7;a grave. O gene adicionado pode ser uma vers&#xE3;o saud&#xE1;vel do gene que &#xE9; mutado no paciente, ou pode ser um gene diferente que inativa ou compensa o gene causador da doen&#xE7;a do paciente. Por exemplo, em pacientes com imunodefici&#xEA;ncia combinada grave (SCID) devido a uma muta&#xE7;&#xE3;o no gene para a enzima adenosina deaminase, uma vers&#xE3;o funcional do gene pode ser inserida. As c&#xE9;lulas do paciente podem ent&#xE3;o produzir a enzima, curando esta doen&#xE7;a potencialmente mortal em alguns casos.
 
 <h4>Inser&#xE7;&#xE3;o Gen&#xE9;tica</h4>
 
 Os genes podem ser introduzidos nas c&#xE9;lulas de um paciente de duas maneiras principais: in vivo&#x2014;diretamente em uma pessoa atrav&#xE9;s da inje&#xE7;&#xE3;o em tecidos espec&#xED;ficos ou na corrente sangu&#xED;nea; e ex vivo&#x2014;em c&#xE9;lulas que foram retiradas do paciente, que s&#xE3;o transplantadas de volta ap&#xF3;s a inser&#xE7;&#xE3;o do gene.
 
 O gene &#xE9; geralmente inserido em um vector&#x2014;muitas vezes um v&#xED;rus que foi modificado para n&#xE3;o causar doen&#xE7;as&#x2014;para que o gene seja introduzido nas c&#xE9;lulas do paciente e entregue ao n&#xFA;cleo. Em alguns casos&#x2014;por exemplo, quando s&#xE3;o usados vectores retrovirais&#x2014;o gene &#xE9; inserido aleatoriamente no genoma da pessoa, levando &#xE0; express&#xE3;o est&#xE1;vel do gene inserido. Em outros&#x2014;como quando s&#xE3;o usados vectores adenovirais&#x2014;o gene n&#xE3;o se integra no genoma do hospedeiro e &#xE9; apenas transientemente expresso. O vector tamb&#xE9;m &#xE9; criado para conter uma regi&#xE3;o de promotor para que o novo gene possa ser transcrito em RNA mensageiro (mRNA). Ent&#xE3;o, usando a pr&#xF3;pria maquinaria da c&#xE9;lula, o mRNA &#xE9; traduzido em prote&#xED;na. O produto proteico&#x2014;como a adenosina deaminase&#x2014;fornece o tratamento para a doen&#xE7;a.
 
 <h4>Ensaios Cl&#xED;nicos</h4>
 
 A terapia gen&#xE9;tica ainda est&#xE1; nos est&#xE1;gios iniciais e traz riscos significativos, em parte porque a integra&#xE7;&#xE3;o e a atividade dos genes inseridos n&#xE3;o podem ser totalmente controladas. Por exemplo, em alguns testes iniciais de terapia gen&#xE9;tica para outra forma de SCID, alguns pacientes desenvolveram cancro, e os ensaios foram interrompidos. No entanto, a tecnologia est&#xE1; a melhorar continuamente, e para doen&#xE7;as graves sem outro tratamento eficaz, os benef&#xED;cios podem superar os riscos. Al&#xE9;m de SCID, outras doen&#xE7;as em que a terapia gen&#xE9;tica foi bem sucedida incluem um tipo de cegueira heredit&#xE1;ria e leucemia de c&#xE9;lulas B resistentes ao tratamento.

Terapia Genética

Gene therapy is a technique where a gene is inserted into a person&#x2019;s cells to prevent or treat a serious disease. The added gene may be a healthy ...

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Education

JoVE Core

Biologia

Genetics

Genética

Biotechnology

Biotecnologia

KEY TERMS AND CONCEPTS

TERMOS-CHAVE E CONCEITOS

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 cases.

Genes can be delivered in two main ways. In vivo gene delivery means the gene is placed directly into the body, often by injection into a specific tissue or into the bloodstream. Ex vivo gene delivery means cells are removed from the patient, given the gene, and then transplanted back.

The gene is usually carried in a vector, which is often a virus that has been changed so it does not cause disease. The vector helps move the gene into the cell and deliver it to the nucleus. Retroviral vectors can insert the gene randomly into the genome, which can lead to stable expression. Adenoviral vectors do not integrate into the host genome, so their gene expression is temporary.

The vector is also engineered with a promoter region. A promoter helps start transcription, so the new gene is copied into messenger RNA, or mRNA. The cell then uses its own machinery to translate the mRNA into protein. That protein, such as adenosine deaminase, is what helps treat the disease.

Gene therapy is still in an early stage and can carry serious risks because inserted genes are hard to control fully. In some early trials for another form of SCID, some patients developed cancer, and the trials were stopped. Even so, the technology continues to improve, and for serious diseases with no other effective treatment, the benefits can outweigh the risks. Successful uses have included inherited blindness and treatment-resistant B-cell leukemia.

Gene Delivery for Serious Diseases

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 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

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.

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

See how gene therapy delivers working genes with viral vectors, in vivo or ex vivo methods, and why the approach can help treat SCID and other diseases.

Gene Delivery for Serious Diseases | JoVE K12

JoVE Core series brings biology to life through over 300 concise and easy-to-understand animated video lessons that explain key concepts in biology, plus more than 150 scientist-in-action videos that show actual research experiments conducted in today’s laboratories.

biology

A coleção JoVE Core dá vida à biologia através de mais de 300 vídeos de animação concisos e fáceis de entender que explicam conceitos-chave de biologia, além de mais de 150 vídeos de Cientistas em ação que mostram experimentos reais em pesquisa realizados nos laboratórios atualmente.

Florida Department of Education

Life Science

SC.912.L.16

SC.912.L.16.10

Molecular Cloning

Molecular cloning is a set of methods, which are used to insert recombinant DNA into a vector - a carrier of DNA molecules that will replicate recombinant ...

An Overview of Genetic Engineering

Genetic engineering &#8211; the process of purposefully altering an organism&#8217;s DNA &#8211; has been used to create powerful research tools and model ...

Recombineering and Gene Targeting

One of the most widely used tools in modern biology is molecular cloning with restriction enzymes, which create compatible ends between DNA fragments that ...

Genome Editing

A well-established technique for modifying specific sequences in the genome is gene targeting by homologous recombination, but this method can be ...

Recombinant DNA

DNA Ligation Reactions

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 ...

Restriction Enzyme Digests

Restriction enzymes or endonucleases recognize and cut DNA at a specific sequence.  These enzymes occur naturally in bacteria as a defense against ...

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further ...

Restriction Enzymes

Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called ...

DNA Agarose Gel Electrophoresis

Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify ...

DNA Gel Electrophoresis

DNA gel electrophoresis is a technique used for the detection and separation of DNA molecules. An electric field is applied to a gel matrix comprised of ...

Separating Protein with SDS-PAGE

Sodium Dodecyl Sulfate Poly-Acrylamide Gel Electrophoresis, or SDS-PAGE, is a widely-used technique for separating mixtures of proteins based on their ...

Plasmid Purification

Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid ...

Gel Purification

Gel purification is used to recover DNA fragments after electrophoretic separation. DNA recovery from an agarose gel includes three basic steps: binding, ...

PCR - Polymerase Chain Reaction

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target ...

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known ...

PCR: The Polymerase Chain Reaction

The polymerase chain reaction, or PCR, is a technique used to amplify DNA through thermocycling &#8211; cyles of temperature changes at fixed time ...

bacterial-transformation-the-heat-shock-method

Bacterial Transformation: The Heat Shock Method

Transformation is the process that occurs when a cell ingests foreign DNA from its surroundings. Transformation can occur in nature in certain types of ...

Bacterial Transformation: Electroporation

The term &#x201C;transformation&#x201D; refers cellular ingestion of foreign DNA.  In nature, transformation can occur in certain types of bacteria.  In ...

An Introduction to Transfection

Transfection is the process of inserting genetic material, such as DNA and double stranded RNA, into mammalian cells. The insertion of DNA into a cell ...

Labeling DNA Probes

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Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside ...

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in ...

An Overview of Gene Expression

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Expression Profiling with Microarrays

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Complementary DNA

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in ...

Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at ...

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their ...

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and ...

Chromatin Immunoprecipitation- ChIP

Chromatin immunoprecipitation, or ChIP, is an antibody-based technique used to identify sites on DNA that bind to transcription factors of interest or ...

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the ...

Synthetic Biology

Synthetic biology is an interdisciplinary science that involves using principles from disciplines such as engineering, molecular biology, cell biology, ...

Genome Annotation and Assembly

The genome refers to all of the genetic material in an organism. It can range from a few million base pairs in microbial cells to several billion base ...

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences ...

The ELISA Method

An enzyme-linked immunosorbent assay (ELISA) is typically performed to detect the presence and/or amount of a target protein of interest within an ...

Transgenic Plants

Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically ...

Transgenic Organisms

Reproductive Cloning

CRISPR

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. ...

Reporter Genes

Reporter genes are a type of&#160;protein-coding gene&#160;that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually ...

In-situ Hybridization

In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled ...

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

Karyotyping

Lampbrush Chromosomes

In 1882, Flemming observed lampbrush chromosomes (LBC) in salamander eggs. Later in 1892, R&#252;ckert observed LBCs in shark egg cells and coined the ...

Polytene Chromosomes

Polytene chromosomes are giant interphase chromosomes with several DNA strands placed side by side. They were discovered in the year 1881 by Balbiani in ...

Euchromatin

The extent of chromatin compaction can be studied by staining chromatin using specific DNA binding dyes. Under the microscope, the dense-compacted regions ...

Heterochromatin

X and Y Chromosomes

Among mammals, the gender of an organism is determined by the sex chromosomes. Humans have two sex chromosomes, X and Y. Every human diploid cell has 22 ...

The Y Chromosome Determines Maleness

The Y chromosome is a sex chromosome&#160;found in several vertebrates and mammals, including humans. In addition to 22 pairs of autosomes, the human ...

The Ratio of X Chromosome to Autosomes

In most organisms, sex is determined by the ratio of X and Y chromosomes. However, in some organisms, such as Drosophila and C.elegans, sex is determined ...

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

Whole Body Regeneration

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Liver Regeneration

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stem-cell-therapy-for-tissue-regeneration

Stem Cell Therapy for Tissue Regeneration

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Methods of Nuclear Reprogramming

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Cloning of Dolly the Sheep

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embryonic-stem-cell-culture-and-differentiation

Embryonic Stem Cell Culture and Differentiation

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Induced Pluripotency

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Analyze scientific and ethical arguments to support the pros and cons of applications of a specific biotechnology technique such as stem cell usage, in vitro fertilization, or genetically modified organisms.
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.

Gene Delivery for Serious Diseases

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