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Learn.genetics.utah --making a transgenic mouse

Source

http://learn.genetics.utah.edu/content/science/transgenic/

Mario caoechhi

Mario R. Capecchi, Ph.D., of the University of Utah, won the 2007 Nobel Prize in Physiology or Medicine. Capecchi shared the prize with Oliver Smithies of University of North Carolina, Chapel Hill, and Sir Martin Evans of Cardiff University in the UK.

The prize recognized Capecchi's pioneering work on "knockout mouse" technology, a gene-targeting technique that has revolutionized genetic and biomedical research, allowing scientists to create animal models for hundreds of human diseases.

As a child, Capecchi wandered homeless in Italy. As a researcher, his first attempts at gene targeting were deemed not ready for funding by the National Institutes of Health. Capecchi is an individual whose personal life proves that while some events are not probable, anything is possible.  Read Mario's story .

During the 1980s, Capecchi devised a way to change or remove any single gene in the mouse genome, creating strains of mice that pass the altered gene from parent to offspring. In the years since, these "transgenic" and "knockout" mice have become commonplace in the laboratory.

Capecchi's pioneering work in gene targeting has taught us much about how the body builds—and rebuilds—itself. He has given scientists worldwide the tools to make important discoveries about human diseases, from cancer to obesity.

And he has raised a key question for the future of human medicine: if we can replace a perfectly good gene with a mutated one, can we also go the other way, replacing problem genes with those that work?

Mario’s lab at work

What makes an arm an arm? Capecchi's research team is working on answering that question using gene targeting. They have systematically "knocked out" a set of genes in mice, called homeotic genes, which govern body patterning during development. For example, one of the lab's most recent genetic discoveries may explain  why we lack spare ribs . Find out more about how homeotic genes work in Genes Determine Body Patterns .

YOUR GOAL: You are studying how a particular gene, named OhNo, might play a role in panic attacks. You want to study mice that are missing this gene. To "knockout" the OhNo gene, you will replace it with a mutated copy that doesn't work.

Here's how:

1. Isolate Stem Cells

Isolate embryonic stem cells that originated from male brown mice with a normal OhNo gene (blue).

2. Add Inactive Gene With Marker

To these cells, add a copy containing a mutated, inactive OhNo gene (red), and a drug resistance marker gene (p3. Similar Genes Naturally Swap

3. Similar Genes Naturally Swap

By mechanisms that are not completely understood yet, similar genes will swap places. The OhNo gene plus drug resistance marker gene is incorporated into the genome, and the normal version is kicked out. This process is called homologous recombination.

ink).

4. Add Drug

Cells that haven't incorporated the inactive OhNo gene don't have the drug resistance marker gene (pink).

Adding the drug kills cells without the marker, leaving you with only cells that have an inactive version of the OhNo gene.

5. Grow Chimeric Mice

By transplanting stem cells that carry the inactive ohNo gene into a white mouse embyro, you'll create what is called a chimera. Chimeras have patches of cells throughout their bodies that grew from white mouse cells and patches that grew from brown stem cells. Some of the cells that have the inactive OhNo gene may develop into reproductive cells.

Chimeras are easy to identify because they have both brown and white patches of fur.

6. Mate Male Chimera

If a male chimera has some reproductive cells (sperm) that originated from the brown stem cells, he will produce some brown offspring when mated with a white female.

7. Test and Breed Brown Offspring

Half of the brown offspring will have a copy of the inactive OhNo gene in all of their cells—including their reproductive cells. These mice have one normal copy of the OhNo gene from their

mother (not shown) and one inactive copy from their father. So half of their reproductive cells will contain a normal copy, and half will contain an inactive copy.

These mice can be identified by performing DNA sequencing in their OhNo genes and then bred with each other.

8. You've Made a Knockout Mouse

One fourth of your resulting offsping will have two copies of the "knocked-out" or inactive OhNo gene. You can now study these mice to determine how lacking the OhNo gene may affect panic attacks.

Supported by a Science Education Partnership Award (SEPA) Grant No. R25RR023288 from the National Center for Research Resources, a component of the NIH. The contents provided here are solely the responsibility of the authors and do not necessarily represent the official views of NIH.

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Questions & Answers

can someone help me with some logarithmic and exponential equations.
Jeffrey Reply
sure. what is your question?
ninjadapaul
20/(×-6^2)
Salomon
okay, so you have 6 raised to the power of 2. what is that part of your answer
ninjadapaul
I don't understand what the A with approx sign and the boxed x mean
ninjadapaul
it think it's written 20/(X-6)^2 so it's 20 divided by X-6 squared
Salomon
I'm not sure why it wrote it the other way
Salomon
I got X =-6
Salomon
ok. so take the square root of both sides, now you have plus or minus the square root of 20= x-6
ninjadapaul
oops. ignore that.
ninjadapaul
so you not have an equal sign anywhere in the original equation?
ninjadapaul
Commplementary angles
Idrissa Reply
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Sherica
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Tamia
what is a good calculator for all algebra; would a Casio fx 260 work with all algebra equations? please name the cheapest, thanks.
Kevin Reply
a perfect square v²+2v+_
Dearan Reply
kkk nice
Abdirahman Reply
algebra 2 Inequalities:If equation 2 = 0 it is an open set?
Kim Reply
or infinite solutions?
Kim
The answer is neither. The function, 2 = 0 cannot exist. Hence, the function is undefined.
Al
y=10×
Embra Reply
if |A| not equal to 0 and order of A is n prove that adj (adj A = |A|
Nancy Reply
rolling four fair dice and getting an even number an all four dice
ramon Reply
Kristine 2*2*2=8
Bridget Reply
Differences Between Laspeyres and Paasche Indices
Emedobi Reply
No. 7x -4y is simplified from 4x + (3y + 3x) -7y
Mary Reply
is it 3×y ?
Joan Reply
J, combine like terms 7x-4y
Bridget Reply
im not good at math so would this help me
Rachael Reply
yes
Asali
I'm not good at math so would you help me
Samantha
what is the problem that i will help you to self with?
Asali
how do you translate this in Algebraic Expressions
linda Reply
Need to simplify the expresin. 3/7 (x+y)-1/7 (x-1)=
Crystal Reply
. After 3 months on a diet, Lisa had lost 12% of her original weight. She lost 21 pounds. What was Lisa's original weight?
Chris Reply
what's the easiest and fastest way to the synthesize AgNP?
Damian Reply
China
Cied
types of nano material
abeetha Reply
I start with an easy one. carbon nanotubes woven into a long filament like a string
Porter
many many of nanotubes
Porter
what is the k.e before it land
Yasmin
what is the function of carbon nanotubes?
Cesar
what is nanomaterials​ and their applications of sensors.
Ramkumar Reply
what is nano technology
Sravani Reply
what is system testing?
AMJAD
preparation of nanomaterial
Victor Reply
Yes, Nanotechnology has a very fast field of applications and their is always something new to do with it...
Himanshu Reply
good afternoon madam
AMJAD
what is system testing
AMJAD
what is the application of nanotechnology?
Stotaw
In this morden time nanotechnology used in many field . 1-Electronics-manufacturad IC ,RAM,MRAM,solar panel etc 2-Helth and Medical-Nanomedicine,Drug Dilivery for cancer treatment etc 3- Atomobile -MEMS, Coating on car etc. and may other field for details you can check at Google
Azam
anybody can imagine what will be happen after 100 years from now in nano tech world
Prasenjit
after 100 year this will be not nanotechnology maybe this technology name will be change . maybe aftet 100 year . we work on electron lable practically about its properties and behaviour by the different instruments
Azam
name doesn't matter , whatever it will be change... I'm taking about effect on circumstances of the microscopic world
Prasenjit
how hard could it be to apply nanotechnology against viral infections such HIV or Ebola?
Damian
silver nanoparticles could handle the job?
Damian
not now but maybe in future only AgNP maybe any other nanomaterials
Azam
can nanotechnology change the direction of the face of the world
Prasenjit Reply
At high concentrations (>0.01 M), the relation between absorptivity coefficient and absorbance is no longer linear. This is due to the electrostatic interactions between the quantum dots in close proximity. If the concentration of the solution is high, another effect that is seen is the scattering of light from the large number of quantum dots. This assumption only works at low concentrations of the analyte. Presence of stray light.
Ali Reply
the Beer law works very well for dilute solutions but fails for very high concentrations. why?
bamidele Reply
how did you get the value of 2000N.What calculations are needed to arrive at it
Smarajit Reply
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Source:  OpenStax, Open genetics. OpenStax CNX. Jan 08, 2015 Download for free at https://legacy.cnx.org/content/col11744/1.3
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