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cmb2000 - essential biomed research > lecture 12 > Flashcards

lecture 12 Flashcards

1
Q

what is genome editing?

A

DNA is inserted, deleted or replaced in the genome of an organism using nucleases:

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2
Q

what is the importance of genome editing?

A

enables specific targeting of sequences within the genome without impacting rest of genome sequence

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3
Q

what is CRISPR/Cas9?

A

enables editing of parts of the genome by removing, adding or altering sections of the DNA sequence

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4
Q

how is CRISPR an adaptive immune regulator?

A

invading DNA recognised and cut by Cas1-Cas2 protein complexes into fragments termed protospacers

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5
Q

where is Cas9/RNA duplex recruited to?

A

complementary sequence on the invading strand of DNA

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6
Q

how does Cas9 prevent infection?

A

cuts DNA strands, creating a double strand break

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7
Q

what is the Cas operon?

A

encodes Cas proteins required for DNA cleavage

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8
Q

what enables Cas9-mediated DNA cleavage?

A

protospacer adjacent motifs (PAM)

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9
Q

are PAM sequence present in the CRISPR locus?

A

no

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10
Q

what 2 components makes gRNA in bacteria?

A

tracrRNA + crRNA

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11
Q

overexpression of Cas9 + gRNA in cells allows…?

A

site direct editing

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12
Q

what are the gRNA expression methods?

A

plasmid, in vitro transcribed RNA

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13
Q

Cas9 expression methods

A

plasmid, in vitro transcribed RNA + recombinant protein

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14
Q

which repair pathways function to repair the DNA?

A

HDR/NHEJ

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15
Q

what is function of NHEJ?

A

enables error-prone DNA repair, key to CRISPR knockout studies

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16
Q

what is function of HDR?

A

enables precise DNA repair, key to CRISPR knock-in studies

17
Q

what happens with CRISPR-mediated gene knock out via NHEJ?

A

normal gene product not expressed

18
Q

what is function of androgen receptor?

A

largely drives progression of prostate cancer

19
Q

current reatment for prostate cancer aims to…?

A

inactivate AR by blocking ligand binding

20
Q

how is AR inactivated?

A

AR gene locus was targeted by CRISPR, gRNA designed to exon 5 of AR with STOP codon

21
Q

individuals with homozygous CCR5delta32 have…?

A

natural resistance to HIV infection

22
Q

individuals heterozygous for CCR5delat32 allele have…?

A

delayed disease progression

cmb2000 - essential biomed research (12 decks)

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