Chick -experiments

Question 1

Is the chick well suited for genetic work?

Answer 1

• NO
• The life cycle is long
• There is no routine protocol for transgenesis or targeted mutagenesis
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Question 2

Overexpression?

Answer 2

• Replicant-competent avian-specifc (RCAS) viruses have been used

Question 3

Electroporation

Answer 3

• Injecting the DNA into the region of interest
• subjecting embryo to low voltage pulses which drives the DNA into the cells
• DNA is negatively charged and so moves towards the anode
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Question 4

How to study embryonic induction?

Answer 4

• Localised treatment of growth factors is done by absorbing the pure factor onto affinity chromoatography beads, which are implanted into the embryo in the desired position.
• Such beads can bind a large amount of growth factor and release it slowly over a number of days mimicing the effects of a signalling centre.
• This is also done in mammalian studies.

Question 5

Loss-of-function?

Answer 5

• This is usually done by dominant negatives
• AMOs are less useful than in zebrafish and xenopus has there are no large blastomeres to inject