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ANBI 470 > M3 L3 > Flashcards

M3 L3 Flashcards

1
Q

what is genome editing

A

genetic engineering that is very specific & localized to a specific location on the DNA

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2
Q

zinc finger nucleases

A

recognize triple repeats & cut at a specific location between recognized sequences
- constructs are made & delivered into embryonic stem cell via viral delivery or electroporation to make transgenic animal

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3
Q

how many basepairs can zinc fingers recognize

A

24

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4
Q

1 advantage & 2 disadvantages of zinc finger nucleases

A

A- can recognize 24bps to be very specific
D- not very triplet has zinc finger nuclease associated with it, and sometimes zinc fingers interact with each other = not efficient

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5
Q

TALENs

A

bind to 1 specific nucleotide & cleave domain

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6
Q

how many bps do TALENs have

A

34

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7
Q

where are TALENs found

A

bacterial pathogens

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8
Q

do TALENs or ZFs have higher specificity?

A

TALENs

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9
Q

what is the newest genome technology

A

CRISPR-Cas9

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10
Q

CRISPR Cas9 meaning

A

CRISPR- clustered regularly interspaced short palindromic repeats

Cas- small clusters of cas (CRISPR-associated system) genes located next to CRISPR sequences

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11
Q

what was CRISPR Cas9 adapted from

A

natural defense mechanisms found in bacteria

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12
Q

how does CRISPR Cas9 work

A

cells that survive infection retain a little piece of DNA in their genome. if similar virus attacks again it will recognize it & destroy it by cutting up the viral genome using cas proteins

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13
Q

what are required for cutting up the foreign DNA

A

cas proteins

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14
Q

CRISPR Cas9 steps

A

1) virus attacks & inserts viral DNA
2) crRNA comes to tracer RNA & matches up with viral genome
3) Cas proteins cut foreign DNA

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15
Q

crRNA

A

CRISPR RNA

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16
Q

tracer RNA function

A

recruits Cas proteins

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17
Q

what interacts with Cas9 protein

A

single guide RNA

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18
Q

Cas steps

A

1) single guide RNA & tracer RNA match up on genome
2) Cas9 & single guide RNA bind
3) complex binds to target sequence & cleaves target DNA
4) cleavage site is repaired by non-homologous end joining or homologous directed repair

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19
Q

what determines if DNA is cleaved in Cas system using non-homologous or homologous repair?

A

homologous- if donor DNA with homologous sequence to the target sequence is present

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20
Q

results of non-homologous end joining vs homologous directed pair in Cas system

A

non-homologous: results possible in gene function disruption
homologous: results in incorporation of new DNA

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21
Q

what nucleotides bind with each other

A

A & T
C & G

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22
Q

3 fates of genome editing

A

1) gene insertion
2) gene correction
3) gene disruption

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23
Q

3 methods of genome editing

A

1) Zinc finger nucleases
2) TALENs
3) CRISPR Cas9

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24
Q

what was the drive of transgenics

A

to study human health

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25
Q

humulin

A

insulin produced by bacteria to mass produce it

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26
Q

1st transgenic livestock species

A

tracy the sheep

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27
Q

tracy the sheep

A

produced recombinant protein -> AAT (alpha 1-antitrypsin) in her milk to treat emphysema & cystic fibrosis

28
Q

transgenic pig rich in omega 3 fatty acids

A

used roundworm gene somatic cell nuclear transfer to create pigs that could produce omega 3

29
Q

what % of children have en egg allergy

A

2

30
Q

hypoallergenic egg

A

used TALENs and CRISPR Cas 9 so chickens born from eggs of transgenic chickens were hypoallergenic

31
Q

PERVs

A

removal of specific proteins in pig genome at same time using CRISPR

32
Q

5 applications of genome transgenic technologies in livestock

A

1) enhanced reproductive performance
2) increased growth rates
3) improved milk production
4) increased resistance to disease
5) improves animal welfare

33
Q

antisense inhibition of polyphenol oxidase

A

prevents discolouration after bruising in potatoes due to less acrylamide

34
Q

T or F: plant transgenics have been around longer than animals

A

T

35
Q

2 negative effects of acrylamide

A

1) nerve damage
2) cancer

36
Q

golden rice

A

beta carotene genes added for increased vitamin A content

37
Q

what can a deficiency in vitamin A cause

A

blindess

38
Q

1st vs 2nd gen golden rice crops

A

1st- not enough transgene
2nd- more beta carotone genes

39
Q

mastitis resistance

A

transfect fibroblasts with transgene that targets cell wall of staph aureus -> somatic cell nuclear transfer -> implant into suurogate -> transgenic cows
- not very successful

40
Q

transgenic chicken

A

when transgene is transcribed, RNA molecule mimics virus to act as molecular decoy so RNA polymerase binds to that instead of actual avian flu virus

41
Q

T or F: transgenic chickens can get infected but they cannot infect other birds

A

T

42
Q

double muscling

A

disruption of myostatin gene using CRISPR Cas9 system
- increases # of muscle fibers & size

43
Q

T or F: FDA approved meat from gene edited pigs for human consumption

A

T

44
Q

T or F: sterile pigs cannot pass transgene to offspring

A

T

45
Q

T or F: slick haired mutation is naturally occurring

A

T

46
Q

CRISPR bacon

A

add in UCP gene into pig genome using CRISPR & somatic cell
- produces leaner meat by burning fat to thermoregulate

47
Q

2 examples of genome editing that enhance animal welfare

A

1) polled mutation
2) slick mutation

48
Q

slick mutation uses what gene

A

prolactin receptor gene

49
Q

slick mutation

A

natural mutation in senepol cattle, produces finer & thinner hair = better regulate body temp in heat = improved animal welfare

50
Q

T or F: slick mutation cattle are approved for consumption

A

T

51
Q

enviro pig

A

genetically modified yorkshire pig that can digest phosphorous more efficiently
- uses phytase to breakdown normally indigestible phosphorous

52
Q

T or F: enviro pigs are still being used

A

F

53
Q

2 types of unintended effects

A

1) off target effects
2) ecological effects

54
Q

off target effects

A

genome editing can lead to unintended mutations which may have unknown implications
- super similar targets = CRISPR for example may target wrong one

55
Q

2 types of ecological effects

A

1) ecosystem disruption -> cannot understand effects on ecosystem
2) gene drives -> can potentially harm biodiversity with transfer of genes across organisms
- mutations can piggy back on gene drives

56
Q

example of gene drive effect

A

mosquitos

57
Q

3 disadvantages of gene drive

A

1) piggy back mutations
2) escape
3) ecological impacts

58
Q

escape

A

can hybridize to another undesired species

59
Q

issues with genome editing

A

revives old social/ethical debates about GMOs, especially related to human editing embryos

60
Q

T or F: if you use a somatic skin fibroblast cell fused to enucleated egg cell, you can create a cloned embryo

A

T

61
Q

T or F: embryonic stem cells from human embryos are not capable of becoming a human being

A

T

62
Q

germline editing regulations in USA, EU & Canada

A

USA- FDA & EPA regulates GMOs
EU- very strict, everything must be labelled
CA- CFIA regulates GMOs
- no germ line editing permitted in canada (cannot work with embryonic stem cells for humans)

63
Q

EPA

A

environmental protection agency
- role for regulation in plants and animals

64
Q

T or F: if you put a polled mutation into an animal that has been bred for polled, they will look the same

A

T

65
Q

what are 4 factors we should consider before moving forward with technological advances?

A

1) applications - therapeutic, but enhancement of personal could be bad (societal standards)
2) equity & access - only for the wealthy?
3) moral, philosophical os religious concerns
- “playing god”
- “disturbing natural processes”
4) balance of risk & benefit

66
Q

Does gene editing impact natural processes?

A

yes, we have been selecting for several years = already interfering

67
Q

disadvantage to not being able to work with embryonic stem cells

A

harder for science to move forward in these areas

ANBI 470 (23 decks)

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