Lab 1

Question 1

Big picture

Answer 1

Does RNAi knockdown of genes that are regulating mitochondrial function alter the toxicity of aggregate formation that models Hunt. Disease

Question 2

exp 1 question

Answer 2

does rnai work in c. elegans?

Question 3

what are we doing in exp 1

Answer 3

worms expressing yfp placed on plates with bacteria containing plasmids l4440 and l4440-yfp. we will measure yfp intensity. we wil compare control with l4440 yfp just to see that rnai works in c elegans.

Question 4

l4440

Answer 4

control vector, will not activate rnai or knock down any proteins

Question 5

l4440-yfp

Answer 5

vector used to knock down yfp. will have yfp sequences in it and bacteria will make dsrna against yfp.

Question 6

exp 2a/2b ques

Answer 6

is polyq toxic? and does knocking down a goi related to mito. function affect polyq toxicity?

Question 7

what’s happening in 2c

Answer 7

same conditions as 2a/2b: knocking down yfp, q35, goi, etc. to see how gene expression changes (seeing gene expression via qpcr)

Question 8

exp 2c ques

Answer 8

how does expression of polyq and/or gene knockdown alter gene expression

Question 9

what’s happening in exp 2a and 2b

Answer 9

worms express q35-yfp (q35 is fused with yfp) or nothing are placed on plates w bacteria containing l4440, l4440-yfp, or l4440-goi. (q35 is the aggregate protein, and that protein will be observed w yfp). in 2a we’ll knock down yfp and q35. in 2b we’re knocking down goi. both will use control.

Question 10

exp 3 ques

Answer 10

does disruption of mictochondrial function activate the cellular stress response?

Question 11

q

Answer 11

glutamine (an amino acid). mutated huntingtin protein has more glutamines than usual. the extended gluatimes lead to protein aggregation

Question 12

q codon

Answer 12

cag

Question 13

proteostasis network

Answer 13

system that ensure all the proteins in a cell are folding/functioning properly

Question 14

proteome

Answer 14

all the proteins in a cell

Question 15

what dictates the function of amino acids?

Answer 15

side chains

Question 16

HSPs

Answer 16

heat shock protein family, chaperones

Question 17

true/false: chaperones use atp to help proteins fold

Answer 17

true

Question 18

proteostasis network consists of

Answer 18

chaperones, proteasomes, and autophagy

Question 19

triggers for folding imbalance

Answer 19

agreggation prone proteins like q35, depletion of chaperones, and aging

Question 20

how to enhance proteostasis

Answer 20

up regulation of chaperones, down regulation of translation, proteasome and autophagy

Question 21

worm types for each exp

Answer 21

1: mus-yfp
2: glp-1 and glp mus-q35