BIO2015: Lecture 6

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  • Created by: LMoney
  • Created on: 12-05-14 13:57
After embryonic cells have been selected, how does one tell if they have survived after injection?
Inject ES cells from brown fur mouse; use blastocyst from black fur mouse - look at the coat colour of the pups
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what is a chimera?
mixture of cells from more than one animal
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what is a gene knockout?
a genetic technique in which one of an organism's genes are made inoperative
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when are gene knockouts used?
in learning about a gene that has been sequenced, but which has an unknown or incompletely known function
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what is an example of a gene knockout that has been studied in mice?
Melanocortin-3 receptor- An important regulator of body fat storage- over 10,000 gene knock-outs described in mice
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what is an example of the application of the pronuclear injection technique?
farm animals as ‘bioreactors'- ‘Pharming’ – pharmaceuticals from farm animals - main application for transgenesis- Potential to replace recombinant bacteria and cell cultures
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what are the advantages of pharming?
Large scale production, low running costs, correct glycosylation and post-translational modification of proteins
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who was 'Tracy'?
first transgenic sheep to produce human protein in milk (1991)
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what was PPL therapeutics a spin off from?
Roslin institute (1987)
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what was Human α1-antitrypsin (AAT) a treatment for?
cystic fibrosis lung disease
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in farm animals, what is the integration efficiency for pronuclear injection?
1-5% (5-40% in mice)- very inefficient
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is it possible to isolate stem cells from farm animals?
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What was the aim of the Dolly research?
To provide an alternative in livestock to mouse embryonic stem cells
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define differentiation
Developmental events between fertilised zygote and mature organism- Specialisation of cells to fulfill specific functions e.g. kidney cell, neurone, etc.
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what was the key development in the dolly research?
Cultured cells induced by starvation to stop growing and enter quiescent (G0) phase of cell cycle - become ‘re-programmable’ when nucleus injected into metaphase II egg
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what was the process of cloning dolly?
full differentiated cell from mammary gland, grown in culture was deprived of nutrients- starvation induced quiescent stage this was fused by electrical shock to an enucleate egg cell to produce egg cytoplasm containing donor cell nucleus
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what did this egg/nucleus fusion mature into?
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how did Dolly die?
Died of a respiratory infection unrelated to cloning, but gave birth to normal lambs, developed premature arthritis (linked to cloning?)
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what is an example of naturally occurring reproductive cloning?
identical twins
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how else can reproductive cloning occur?
embryo splitting
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is cloning by nuclear transfer transgenesis or not?
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how can nuclear transfer be applied to livestock?
genetic improvement- allows rapid introduction of genes from elite animals- can also be used for conservation of endangered breeds of animals
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what is the impact of nuclear transfer on making transgenic farm animals?
nuclear transfer allows mouse stem cell methods to be used, providing precise gene targeting – second generation of transgenic livestock More sophisticated approaches possible than first generation of transgenic livestock by pronuclear injection
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what is a chimera?


mixture of cells from more than one animal

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what is a gene knockout?


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when are gene knockouts used?


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what is an example of a gene knockout that has been studied in mice?


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