L6: Functional Genomes Flashcards

1
Q

What are strategies to explore genes and function

A

CRISPR => site specific
RNAi => specific
Transposon mediated mutagenesis => non specific target

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2
Q

Why use a transposon approach

A
  • elements can be mobilised or immobolised on demand
  • needs few transformants to generate high number of org w/ transposed elements at varying locations
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3
Q

How do you make primary Ds Transposon lines

A
  • co transformation (bombardment) of immature embyros with plasmids with Ds elements
  • leading to Ac transposase initated by primary transposition
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4
Q

What does the two component transposon system require

A
  • transposases
  • jump
  • landing
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5
Q

Give the an example of transgenic organisms

A

PAPAYA
* 1970: ringspot virus was found
* 91: developed transgenic w/ virus resistance
* 98: approved, ‘vaccinated’ with protein coat to induce SAR and make them resistant to virus

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6
Q

What is gene editing

A
  • targetted alteration of DNA sequences
    CRISPR
    clustered regularly interspaced short palindromic repeats
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7
Q

History of gene editing

A

1996: Zinc finger nucleases
2003: used mega nucleases to edit genes
2010: Talen
2012-23: CRISPR Cas

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8
Q

What is the different in cas9 mechs

A

nucleotide deletion => mutation with DNA repair NHEJ

nucleotide addtion: mutation with DNA tweaking (HDR)

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9
Q

Discuss gene regulation by USDA

A

Deletions: the change to the plant is solely a genetic deletion of any size.

  • Single base pair substitutions: the change to the plant is a single base pair substitution.
  • Insertions from compatible plant relatives: the change to the plant solely introduces nucleic acid sequences from a compatible relative that could otherwise cross with the recipient organism and produce viable progeny through
    traditional breeding.
  • Complete null segregants: offspring of a genetically engineered
    plant that does not retain the change of its parent.
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