Lecture 17: Genetic Manipulation in ES Cells

Podcast

Play an AI-generated podcast conversation about this lesson

Questions and Answers

Which characteristic is essential for embryonic stem (ES) cells?

Incapable of culturing in a laboratory environment

Ability to undergo self-renewal while remaining undifferentiated (correct)

Ability to differentiate into a single cell type only

Formation of specialized cells without epigenetic changes

What is the primary source of embryonic stem cells in a developing embryo?

The inner cell mass of the blastocyst (correct)

The blastocoel

The placenta

The trophoblast layer

What distinguishes knockout mice from knock-in mice?

Knockout mice have specific genes deleted, whereas knock-in mice have specific genes inserted (correct)

Knockout mice have added genes while knock-in mice do not

Knock-in mice are generated from embryonic stem cells only

Both types of mice have identical genetic modifications

What is a significant outcome of epigenetic reprogramming in pluripotent stem cells?

It enhances the ability to undergo specific differentiation Signup and view all the answers

What technique can be utilized to introduce DNA into embryonic stem cells?

Electroporation Signup and view all the answers

How do CRISPR-mediated genome editing and ES cells interact?

CRISPR is used to introduce mutations into ES cells for research Signup and view all the answers

What is the role of selectable markers in genetically modified embryonic stem cells?

To enable identification and selection of successfully modified cells Signup and view all the answers

What is a critical difference between transgenic mice and knockout mice?

Transgenic mice are modified to express foreign genes, while knockout mice have genes deleted Signup and view all the answers

What is the primary purpose of using CRISPR in conventional gene targeting in mice?

To directly modify genes in embryonic stem cells Signup and view all the answers

In the context of gene editing, why is the microinjection of sgRNA and Cas9 protein complex into a mouse zygote significant?

It allows for faster creation of knockout mice in a single step. Signup and view all the answers

What characterizes knockout mice primarily examined in research?

Homozygous deletion of specific genes. Signup and view all the answers

What is a key distinction between knock-in and knockout mice?

Knock-in mice involve the introduction of a specific mutation at a genetic locus. Signup and view all the answers

What role does the male pronucleus play in creating transgenic mice?

It serves as the primary site for random integration of injected DNA. Signup and view all the answers

Which of the following statements about the phenotypes of heterozygous mutants is most accurate?

They may express traits due to haploinsufficiency. Signup and view all the answers

In which area can CRISPR experiments in embryonic stem (ES) cells be applied?

In vitro differentiation and development of organoids. Signup and view all the answers

What is the outcome of the example mention of a CRISPR knock-in mouse line?

A point mutation altering tyrosine 142 to alanine in H2AX. Signup and view all the answers

What defines totipotency in a cell?

The capacity to produce all differentiated cell types in an organism Signup and view all the answers

What is a consequence of the reprogramming process involved in the induction of iPS cells?

Loss of epigenetic modifications Signup and view all the answers

Which of the following statements best describes embryonic stem (ES) cells?

They can give rise to all cell types found in an adult organism Signup and view all the answers

What role do fibroblast feeder cells play in culturing ES cells?

They provide a non-proliferating layer to support ES cell growth Signup and view all the answers

What is the result when altered ES cells are injected into a wild-type blastocyst?

A chimera will develop with contributions from both ES cells and host cells Signup and view all the answers

Which of the following statements about chimera mice is accurate?

They may possess a mix of germ cells from both origins Signup and view all the answers

What happens to the embryonic cells during the cleavage divisions of a mammalian embryo up to the eight-cell stage?

Each cell remains totipotent and capable of forming a complete organism Signup and view all the answers

What distinguishes pluripotent cells from totipotent cells?

Pluripotent cells lack the ability to form an entire organism Signup and view all the answers

Study Notes

Lecture 17: Genetic Manipulation in Embryonic Stem Cells and Animals

Totipotency, pluripotency, and germline biology are discussed.
Blastocyst development (days 3-5): Crucial video for understanding the process. (Link provided)
Embryonic stem (ES) cells: Defined as cells derived from the inner cell mass of a blastocyst, capable of differentiating into all cell types in the body. They can be cultured and used to generate genetically modified animals.
Genetically modified animal models: ES cells enable researchers to create animals with specific genetic alterations for studying various biological processes.
CRISPR experiments in ES cells: Gene editing tools like CRISPR are used to modify the genome of ES cells.
Introduction to mouse genetics: Mouse genetics is important in researching genetic manipulations (knockouts, knock-ins, transgenics).
Embryonic stem cell features: ES cells have the capacity for self-renewal and differentiation.
CRISPR-mediated genome editing in ES cells: Described as a technique where DNA sequences are precisely altered using CRISPR.
CRISPR-mediated genome editing in creating genetically modified animal models: Explains how gene editing is used to create genetically modified animals.
Knockout, knock-in, versus transgenic mice: Different genetic manipulation strategies that produce genetically altered mice are differentiated.
Making pluripotent cells (iPS cells): Technique of inducing pluripotency by forcing expression of particular transcription factors.
Totipotency: Ability of a single cell to develop into a complete adult organism. ES cells exhibit pluripotency, the capacity to differentiate into all cell types but not a whole organism.
ES cell isolation and culture: Techniques used for isolating and maintaining human or mouse ES cells. Includes a layer of fibroblast feeder cells to support their growth.
Chimera mice: Embryos formed by introducing genetically modified ES cells into the blastocyst of a wild-type mouse, resulting in animals with a mix of genetically modified and normal cells.
Conventional gene targeting in mice: Using ES cells to create targeted alterations in the genome of mice.
CRISPR experiments in ES cells: Involves similar strategies used in other cell types, with many applications in stem cell research (in vitro studies, differentiation, organoids, and therapies).
Introduction to mouse genetics: Discusses different types of genetic modifications to specific genes, such as knockouts, knock-ins, and transgenics. Highlights how these techniques are used to create genetically modified mice.
Viral infection/transfection: Methods for introducing DNA into ES cells (and other cells in similar experiments).
Selectable markers: Used to identify and select cells with the desired genetic modification.
Specific examples: The use of CRISPR and targeted gene editing in various applications are discussed including the H2AX gene.

Studying That Suits You

Use AI to generate personalized quizzes and flashcards to suit your learning preferences.

Description

Explore the fascinating world of genetic manipulation in embryonic stem cells and animals. This lecture covers key concepts like totipotency, pluripotency, and the impact of CRISPR on gene editing. Discover how genetically modified animal models aid in biological research.