Name: crispr gene editing basics
Uploaded: 2021-04-07T12:12:00.000Z
Duration: 1 min 59 s
Channel: JoVE

K12 Standards

north-carolina-k-12-science-standard-course-of-study

NC K-12 Science Standard Course of Study

9-12

Biology

heredity-inheritance-and-variation-of-traits

Heredity- Inheritance and Variation of Traits

LS.Bio.8.2

CRISPR

CRISPR-Cas9 시스템은 유전자 편집 기술로서그 의미는 무리진, 규칙 이격이고짧고, 회문 반복 및 CRISPR 관련 단백질 9이다박테리아에서 처음 관찰된 CRISPR-Cas9는바이러스에 맞서는 방어 수단이다외부 바이러스 DNA가 박테리아에 들어오면더 작은 조각으로 나뉘고한 부분에 삽입이 되는데그 부분은 CRISPR 위치라는 박테리아 유전체이다이 부분이 전사될 때, 그 결과물은tracrRNA라는 더 작은 RNA와 연계되는데그 분자에 대한 Cas9 단백질과 RNAse의방향을 잡는데 도움을 주고, 후자는전사물을 잘라준다최종 결과는 여러 개의 복합물인데그 각각은 Cas9 단백질, tracrRNA그리고 DNA 위치에서 나온 CRISPR-RNA이다이 구조물에 있는 CRISPR-RNA는Cas9을 인지하고 바이러스 DNA로 안내하는데그리고 나서 잘리고 파괴가 된다과학자들은 CRISPR-Cas9을 이용하여tracrRNA와 crRNA를 모방하는RNA 분자를 합성하는데이 분자는 문제의 유전자를 표적으로 잡는다이를테면, 두 개의 그러한 안내 RNA가Cas9을 가진 세포에 도입되고둘 다 동일한 유전자를 표적으로 잡으면, 서열이 잘려진다이 표적 지역이 제거되면잘린 끝은 다시 연결되고세포에 대한 영향을 관찰할 수 있다그래서, CRISPR-Cas9 시스템은박테리아 기제로부터 수정이 되고여러 유전자 편집 기술로 이용될 수 있다

Thurtle-Schmidt, Deborah M., and Te‐Wen Lo. "최첨단의 분자생물학: 학부생을 위한 CRISPR/CAS9 유전자 편집에 대한 검토." Biochemistry and Molecular Biology Education 46, no. 2 (2018): 195–205. [<a href="https://doi.org/10.1002/bmb.21108" target="__blank">소스</a>]

Lander, Eric S. "The Heroes of CRISPR." Cell 164, no. 1 (2016년 1월 14일): 18–28. [<a href="https://doi.org/10.1016/j.cell.2015.12.041" target="__blank">출처</a>]

크리스퍼

유전체 편집(genome editing) 기술은 과학자들이 특정 유전체 위치에 유전 물질을 추가, 제거 또는 재배치해서 유기체의 DNA를 수정할 수 있게 해줍니다. 이런 유형의 기술은 잠재적으로 혈우병(hemophilia)과 겸상적혈구 빈혈(sickle cell anemia)과 같은 유전 질환을 치료하는데 사용될 수 있습니다. 유전 질환에 대한 안전하고 효과적인 치료법을 이끌어 낼 가능성이 있고, 또한 널리 사용되고 있는 DNA 편집 도구 중 하나는 CRISPR/Cas9(크리스퍼/캐스나인) 시스템입니다. CRISPR/Cas9은 Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein 9의 약자입니다. 기본 CRISPR/Cas9 시스템은 Cas9 핵산내부가수분해효소(endonuclease)와 Cas9을 표적(target) DNA로 유도하는 작은 RNA로 구성됩니다.
 
 <h4>기원</h4>
 
 CRISPR 서열은 박테리아에서 처음 관찰되었고 나중에 고세균(archaea)에서도 확인되었습니다. 연구자들은 CRISPR/Cas9 시스템이 침입하는 바이러스에 대한 적응면역(adaptive immunity; 후천면역) 방어 기능을 제공한다는 것을 발견했습니다. 여러 박테리아와 대부분의 고세균은 바이러스 DNA의 짧은 염기서열을 포착하여 바이러스 DNA 조각의 모음(library), 즉 CRISPR 배열(CRISPR array)을 만듭니다. 원핵생물(prokaryote)이 같은 바이러스나 바이러스의 종류에 다시 노출될 때, CRISPR 배열은 바이러스 침입자를 인지하는 데 도움이 되는 작은 RNA 조각을 전사하고, 그 후에 Cas9 또는 유사한 핵산내부가수분해효소로 바이러스 DNA를 파괴하는 데 사용됩니다.
 
 <h4>CRISPR/Cas9 기술의 사용</h4>
 
 CRISPR/Cas9은 DNA를 제거하고 새로운 DNA 서열을 그 자리에 삽입하기 위해 실험실에서 흔히 사용됩니다. 이를 위해 먼저 가이드 RNA(guide RNA)라고 불리는 작은 RNA 조각을 만들어야 하는데, 여기에 유전체 DNA의 특정 표적 서열에 결합하는 가이드 서열(guide sequence)이라고 불리는 짧은 서열도 포함시켜야 합니다. 가이드 RNA는 Cas9(또는 Cpf1과 같은 다른 핵산내부가수분해효소)과도 결합할 수 있습니다. 가이드 RNA와 Cas9 단백질은 관심 세포(cell of interest)에 투입되며, 여기서 가이드 RNA는 표적 DNA 서열을 식별하고 Cas9는 그것을 절단합니다.
 
 그다음 세포의 내부 체계는 무작위 뉴클레오타이드(nucleotide)를 삽입 또는 삭제해 손상된 가닥을 수리하여 표적 유전자를 비활성화합니다. 한편 맞춤형 DNA 서열이 가이드 RNA와 Cas9과 함께 세포에 도입될 수 있으며, 이 맞춤형 DNA 서열이 손상 복구 체계의 주형(template)으로 사용되어 잘려 나간 서열을 대체합니다. 이것은 연구자들이 유전자제거를 통해 유전자의 효과를 연구하거나 질병을 치료하기 위해 돌연변이 유전자를 정상적인 사본으로 대체하는 매우 효과적인 방법입니다.
 
 <h4>윤리적, 도입 가능성 고려 사항</h4>
 
 CRISPR/Cas9 시스템이 유전자 조작에 활용될 수 있는 점 때문에, (특히 배아 편집과 관련하여) CRISPR/Cas9 사용에 대한 큰 논쟁이 있었습니다. 중국의 한 과학자는 HIV 감염에 관련된 유전자를 비활성화하기 위해 CRISPR 기술을 사용하여 유전체 편집 아기를 만들었다고 주장했습니다. 이는 절차의 윤리적, 안전적 고려에 대해 우려하는 과학자들로부터 세계적인 항의를 이끌어 냈습니다. 많은 사람들은 이 조치가 시기상조라고 주장했고, 다른 사람들은 표적 밖의 유전체에 미치는 영향에 대한 우려를 표명했습니다. CRISPR/Cas9 시스템의 생명공학 응용 가능성은 크지만, 사용으로 인해 발생할 수 있는 미래의 문제를 고려하는 것이 중요합니다.

게놈 편집 기술을 통해 과학자들은 특정 지역에서 유전 물질의 추가, 제거 또는 재배열을 통해 유기체의 DNA를 수정할 수 있습니다.

Education

교육

JoVE Core

Molecular Biology

분자생물학

Unit 2

제2단원

Analyzing Gene Expression and Function

유전자 발현 및 기능 분석

KEY TERMS AND CONCEPTS

주요 용어 및 개념

crispr gene editing basics

CRISPR is a gene editing tool that helps scientists make targeted changes to DNA. It uses a guide system to find a chosen sequence and a cutting protein to act at that spot.

This method is based on a natural defense system found in bacteria. In high school biology, it is often studied as a way to see how genes can be edited with precision.

CRISPR is widely used in molecular biology research. It helps scientists explore how genes function and how DNA changes can affect living things.

CRISPR Gene Editing Basics

in vitro mutagenesis

In Vitro Mutagenesis for Gene Analysis

In vitro mutagenesis is a lab method used to change DNA and study gene function. It helps scientists test how specific DNA changes affect a gene’s activity and the traits it controls. A common approach is to make a known change in a DNA sequence and then examine the result. This can be done by deleting, inserting, or altering selected bases in a gene. The method is useful for linking a DNA change to a biological effect. Researchers use in vitro mutagenesis in molecular biology, genetics, and...

genetic screens

Using Genetic Screens to Find Disease Genes

Genetic screens are tools used to find genes and mutations linked to specific traits. They are also used to identify people or groups at risk for genetic diseases. That can support early intervention, targeted therapy, and reproductive choices. Forward genetic screens, also called classical screens, start with random DNA changes in an organism. Researchers may use radiation, mutagens, or added bases to create mutations that lead to visible phenotype changes. The mutant is then inbred so...

test cross

Test Cross in Genetic Inheritance

A test cross is a genetics tool used to determine the genotype of an organism with a dominant trait. It helps reveal whether the dominant individual is homozygous or heterozygous by crossing it with a recessive individual. The outcome of the cross shows the hidden genotype. If any offspring show the recessive trait, the dominant parent must carry one recessive allele. If all offspring show the dominant trait, the dominant parent is likely homozygous dominant. Test crosses are common in basic...

complementation tests

Using Crosses to Group Mutations

A complementation test uses a simple cross to find out whether two mutations are in the same gene or in different genes. It helps show how mutations are arranged on chromosomes. Edward Lewis first performed this test in the 1940s while studying fruit flies. To do the test, organisms heterozygous for different mutations are crossed pairwise in all combinations. If the mutations are in different genes, they can complement each other. In that case, each mutation provides the missing genetic...

single nucleotide polymorphisms-snps

SNPs as Genetic Markers and Variants

Single nucleotide polymorphisms, or SNPs, are small DNA changes at one specific genomic position. They are the most common type of sequence variation in the human genome. A point mutation counts as an SNP when it appears in more than 1% of the population.

bacterial transformation

Bacterial Transformation in the Lab

Bacterial transformation is a lab method used to move new DNA into bacteria. Scientists use it to change how bacteria behave or to help them carry a gene of interest. In high school biology, this process is often shown with plasmids, which are small circular DNA molecules. During transformation, bacteria take up DNA from their surroundings. The new DNA can then be used by the cell, depending on the experiment. This technique is an important part of genetic engineering and is widely used in...

transgenic organisms

Making Transgenic Organisms

Transgenic organisms are living things with DNA added from another organism. A transgene is the inserted gene. Scientists use this method to change an organism’s traits and to study how genes work. To make a transgenic organism, researchers first identify the gene they want to add. They then place that gene into the target organism’s DNA. Once the gene is integrated, the organism can pass it to new cells as it grows. This approach is useful in biology because it helps scientists analyze gene...

reproductive cloning

Cloning an Organism by Reproduction

Reproductive cloning creates a new organism that is genetically the same as the donor. It is different from cloning genes or cells because the goal is to make a whole living individual. This process usually relies on somatic cell nuclear transfer. In this method, the nucleus from a body cell is placed into an egg cell that has had its own nucleus removed. The egg can then develop into an embryo with DNA from the donor cell. Reproductive cloning is used to produce a copy of an organism, such...

crispr

CRISPR is a gene editing tool that helps scientists make targeted changes to DNA. It uses a guide system to find a chosen sequence and a cutting protein to act at that spot. This method is based on a natural defense system found in bacteria. In high school biology, it is often studied as a way to see how genes can be edited with precision. CRISPR is widely used in molecular biology research. It helps scientists explore how genes function and how DNA changes can affect living things.

experimental rnai

RNAi for Gene Silencing and Discovery

RNA interference (RNAi) is a gene-silencing process used to reduce gene expression. It can block transcription or trigger RNA degradation. Scientists first discovered this mechanism in plants in 1998 through Andrew Fire and Craig Mello. RNAi has since been found in many eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. Researchers use it as a practical tool to study gene function. It can show what a gene does without permanently changing the organism’s DNA. <br...

reporter genes

Reporter Genes in Cells and Plasmids

Reporter genes are protein-coding genes used to track gene activity in cells. They are often tagged to a gene of interest so scientists can see whether that gene is present or active. When the tagged genes are expressed together, reporter genes can produce visible signals such as fluorescence or luminescence. These signals help show the presence or absence of a gene of interest in an organism. They can also reveal the pattern of gene expression or the location of a DNA segment or protein...

in-situ hybridization

ISH Probes for DNA and RNA Detection

In situ hybridization (ISH) uses labeled probes to detect and localize specific DNA or RNA molecules in cells, tissues, or tissue sections. The probe is a complementary strand of DNA or RNA that binds to a matching nucleotide sequence in the sample. Since its first use in 1969 for nucleic acid studies, ISH has become an important tool in research and in clinical diagnosis. Several probe types can be used in ISH. These include single-stranded DNA probes, double-stranded DNA probes, antisense...

chromatin immunoprecipitation- chip

ChIP Methods for DNA-Protein Binding

Chromatin immunoprecipitation, or ChIP, is an antibody-based method for finding where proteins bind to DNA. It is used to identify DNA sites bound by transcription factors or histone proteins. ChIP also helps show which histone changes are present, including acetylation, phosphorylation, and methylation. ChIP is commonly divided into two forms: X-ChIP and N-ChIP. X-ChIP first cross-links proteins to DNA inside the cell, then breaks the DNA into pieces by sonication. The protein-DNA complexes...

synthetic biology

Synthetic Biology in Crops and Cells

Synthetic biology combines engineering, molecular biology, cell biology, and systems biology to redesign living systems. Scientists use it to remodel organisms from nature or to build new synthetic organisms. These tools can support protein or enzyme production, bioremediation, the production of useful macromolecules, and the addition of helpful traits to crops. One well-known example is golden rice. This genetically modified rice plant makes grains rich in β-carotene, which is a precursor of...

ribosome profiling

Ribosome Profiling for Translation Snapshot

Ribosome profiling is a deep sequencing method that gives a snapshot of active translation in a cell. It sequences the mRNAs protected by ribosomes. This shows which messages are being read to make proteins at a specific moment. The method has several uses in biology. It can track translation in a living organ or tissue type. It also helps measure new protein synthesis levels. Researchers use it to find translated open reading frames, which are stretches of RNA that can make protein. It can...

transgenic plants

Transgenic Plants for Better Crops

Transgenic plants are crops that have been changed with recombinant DNA technology, also called transgenesis. This method adds a foreign gene or removes a harmful one from an organism. The result is a genetically modified organism. The first transgenic plant was a tobacco plant made in 1983, and it was resistant to tobacco mosaic virus. Since then, transgenic plants have been developed for many uses in agriculture, horticulture, and ornamental plants. Some are designed to resist pests or...

gene therapy

Gene Therapy for Disease Treatment

Gene therapy is a treatment approach that changes gene activity to help address disease. It focuses on the genetic cause of a problem rather than only easing symptoms. In this way, it is used in disease intervention. One common strategy is to replace a faulty gene with a normal copy. Another strategy is to inactivate a gene that is causing harm. Gene therapy can also introduce a new gene to help cells work more normally.

epistasis analysis

Gene Interaction Maps in Epistasis

Gene interaction maps in epistasis show how different genes work together to shape a phenotype. In epistasis, the interaction of genes or alleles at different locations affects the final trait. This often happens when one gene masks or interferes with the expression of another gene. Epistasis is common when genes are part of the same biochemical pathway. In that case, the expression of one gene can depend on a gene product made earlier in the pathway. Genes that mask or inhibit other genes in...

genome-wide association studies-gwas

Using SNP Patterns to Link Disease Risk

Genome-wide association studies, or GWAS, look for common SNPs, or single nucleotide polymorphisms, that may be linked to disease. Researchers compare how often a SNP appears in people with a disease and in people without it. When a SNP is seen more often in the disease group, it is considered associated with that disease. GWAS uses chi-square analysis to test the chance that an allele is linked to the disease. This approach does not require scientists to first find the target gene behind the...

key terms and concepts

Explore CRISPR as a gene editing tool, its bacterial origins, and how it is used in molecular biology to study DNA and gene function.

CRISPR Gene Editing Basics | JoVE K12

molecular biology

JoVE Core 분자생물학은 명확하고 간결한 애니메이션을 통해 분자 단계에서 일어나는 세포 과정을 설명합니다.또한, 과학자가 직접 실험을 진행하는 실사 영상을 통해 분자생물학 개념의 응용 사례를 보여줍니다.

Tennessee Academic Standards

High School

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

Links Among Engineering, Technology, Science, and Society

BIO1.ETS2.1

Virginia  Standards of Learning

HIgh School

BIO.5

BIO.5.e

Arizona Science Standards

Life Science

HS.L3

HS.L3U3.26

Alabama Course of Study Science

from-molecules-to-organisms-structures-and-processes

From Molecules to Organisms: Structures and Processes

9-12.B.1.2.c

Florida Department of Education

SC.912.L.16

SC.912.L.16.7

myJoVE Corporation

What is Meiosis?

Meiosis is the process by which diploid cells divide to produce haploid daughter cells. In humans, each diploid cell contains 46 chromosomes, half from ...

Meiosis I

Meiosis II

Meiosis vs. Mitosis

Crossing Over

Crossing over is the exchange of genetic information between homologous chromosomes during prophase I of meiosis I. Genetic recombination gives rise to ...

Nondisjunction

Nondisjunction is the failure of homologous chromosomes or sister chromatids to separate correctly and move to the opposite poles of the cells. This ...

Spermatogenesis

Spermatogenesis is the process by which haploid sperm cells are produced in the male testes. It starts with stem cells located close to the outer rim of ...

Oogenesis

In human women, oogenesis produces one mature egg cell or ovum for every precursor cell that enters meiosis. This process differs in two unique ways from ...

Fertilization

During fertilization, an egg and sperm cell fuse to create a new diploid structure. In humans, the process occurs once the egg has been released from the ...

Cleavage and Blastulation

After a large-single-celled zygote is produced via fertilization, the process of cleavage occurs while zygotes travel through the uterine tube. Cleavage ...

Gastrulation

Gastrulation establishes the three primary tissues of an embryo: the ectoderm, mesoderm, and endoderm. This developmental process relies on a series of ...

Neurulation

Neurulation is the embryological process which forms the precursors of the central nervous system and occurs after gastrulation has established the three ...

Cell Migration

Cell migration, the process by which cells move from one location to another, is essential for the proper development and viability of organisms ...

Determination

During embryogenesis, cells become progressively committed to different fates through a two-step process: specification followed by determination. ...

Use models to explain how DNA is passed from parents to offspring through the processes of meiosis and fertilization in sexual reproduction.

LS.Bio.6.1

Mutations

Mutations are changes in the sequence of DNA. These changes can occur spontaneously or they can be induced by exposure to environmental factors. Mutations ...

genome-size-and-the-evolution-of-new-genes

Genome Size and the Evolution of New Genes

types-of-genetic-transfer-between-organisms

Types of Genetic Transfer Between Organisms

Base-pairing and DNA Repair

The DNA Replication Fork

Lagging Strand Synthesis

The Replisome

Proofreading

Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template ...

Replication in Prokaryotes

DNA replication has three main steps: initiation, elongation, and termination. Replication in prokaryotes begins when initiator proteins bind to the ...

Replication in Eukaryotes

In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before ...

Overview of DNA Repair

Base Excision Repair

Nucleotide Excision Repair

DNA Distortion and Damage
Cells are regularly exposed to mutagens&#8212;factors in the environment that can damage DNA and generate mutations. UV ...

Mismatch Repair

Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the ...

Fixing Double-strand Breaks

Homologous Recombination

Gene Conversion

Restarting Stalled Replication Forks

Construct an explanation to summarize how inheritable genetic variations may result from: new genetic combinations in meiosis, mutations during replication, or mutations caused by environmental factors.

LS.Bio.6.2

An Overview of Genetic Analysis

An organism&#8217;s physical traits, or phenotype, are a product of its genotype, which is the combination of alleles (gene variants) inherited from its ...

Genetic Crosses

To dissect genetic processes or create organisms with novel suites of traits, scientists can perform genetic crosses, or the purposeful mating of two ...

Punnett Squares

Monohybrid Crosses

Dihybrid Crosses

Trihybrid Crosses

Trihybrid Crosses
Some of Mendel&#8217;s crosses examined three pairs of contrasting characteristics. Such a cross is called a trihybrid cross. A ...

Law of Independent Assortment

Chi-square Analysis

The chi-square test is a statistical hypothesis test. It is used to check whether there is a significant difference between an expected value and an ...

Pedigree Analysis

Multiple Allele Traits

Incomplete Dominance

Gregor Mendel&#39;s work (1822 - 1884) was primarily focused on pea plants. Through his initial experiments, he determined that every gene in a diploid ...

Lethal Alleles

Agouti: A Lethal Allele
Lucien Cu&#233;not discovered lethal alleles in 1905 while studying the inheritance of coat color in mice. The agouti gene is ...

Polygenic Traits

background-and-environment-affect-phenotype

Background and Environment Affect Phenotype

Although the genetic makeup of an organism plays a major role in determining the phenotype, there are also several environmental factors, such as ...

Use mathematics and computational thinking to predict the variation and distribution of expressed traits based on: Mendelian inheritance, co-dominance, incomplete dominance, multiple alleles, and sex-linked inheritance.

LS.Bio.7.1

Overview

A pedigree is a diagram displaying a family&#x2019;s history of a trait. Analyzing pedigrees can reveal (1) whether a trait is dominant or ...

Probability Laws

Overview

The probability of inheriting a trait can be calculated using the sum and product rules. The sum rule is used to calculate the probability of ...

The Concept of Multiple Allelism

Multiple allelism describes genes that exist in three or more allelic forms. Although diploid organisms, like humans, ...

Analyze and interpret data to explain how polygenic traits result in a wide range of phenotypes.

LS.Bio.7.2

Pleiotropy

Pleiotropy is the phenomenon in which a single gene impacts multiple, seemingly unrelated phenotypic traits. For example, defects in the SOX10 gene cause ...

Nature and Nurture

Many human characteristics, like height, are shaped by both nature&mdash;in other words, by our genes&mdash;and by nurture, or our environment. For ...

Construct an explanation to summarize how traits result from interactions of genetic factors (multiple genes and/or alleles) and environmental factors.

LS.Bio.7.3

Understand genetic mechanisms for variation.

CRISPR Gene Editing Basics

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