Genome Surveillance

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Genome Surveillance

• ...epigenetic control of gene expression by RNA
  processing is considered a component of normal
  development,
• defense against viruses,
• control of transposable elements,
• regulate gene expression?

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New Phenomenon

• Grind em, find em.

Nature makes em, we breaks em.
insight
Genomics
Broader Understanding
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Nature, 404, pp 293
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Drosophila S2 Cells(cell tissue cultures)

• ...loss-of-function phenotypes?
• targeted degradation of exogenous RNA,
• targeted degradation of endogenous RNA,
• RNAi () activity, RNAi(-) activity.

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Figure 1a.
What does this figure establish?

• Transient Expression of b-galactosidase?
• Transfection?
• CD8 ds RNA?
• lacZ dsRNA?

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Figure 1b.
What does this figure establish?

• Endogenous Gene Silencing?
• Count?
• PI?

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Figure 1b.
What does this figure establish?

• Endogenous Gene Silencing?
• Count?
• PI?

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Figure 1c.
What does this figure illustrate?

• Northern blot?
• Whats bound to the membrane?
• Whats labelled?

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Figure 2a.
What does this figure establish?

• Cellular lysates, from cells treated how?
• Synthetic mRNA transcripts?
• Whats bound to the membrane?
• Whats labelled?

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Figure 2b.
What does this figure show?

• Whats the lysate?
• Whats the target?
• Whats bound to the membrane?
• Whats labelled?

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Figure 2c.
What does this figure show?

• Whats the lysate?
• Whats the target?
• Whats bound to the membrane?
• Whats labelled?

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Figure 3.
What does this figure establish?

• Whats a Ca-dependent nuclease?
• What does microccocal nuclease do?
• What does EGTA do?
• Why assay DNAse I activilty in vitro?

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Figure 4a.
What does this figure establish?

• Whats labelled?
• why the sense strand?
• Whats bound to the membrane?
• what steps did the RNA and associated molecules
  go though prior to gel electrophoresis? Why?
• Why is there so much signal?

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Figure 4b.
What does this figure establish?

• Whats labelled?
• Whats bound to the membrane?
• Whats different in each lane?

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