Lecture 7

Question 1

Any organism in which foreign DNA is introduced and then becomes a stable genome is also known as _______-ic

Answer 1

transgen

Question 2

When a transgene is expressed, cells marked with a neutral enzyme (promote/degrade) similar cells.

Answer 2

promote

Question 3

When a transgene is expressed, the alternate outcome can also be that it ________ the protein. (think Alzheimer’s)

Answer 3

overexpresses

Question 4

Transgenesis helps us to find out ______ in the KO animal.

Answer 4

issues

Question 5

Pronuclear injection is used to _____ a _____ gene by injecting foreign DNA into a fertilized egg and seeing how it affects brain function.

Answer 5

modify ; desired

Question 6

The five steps in transgenesis are:

1. injecting ______
2. allowing ____ to integrate with _____ gene
3. observe ___ or ____ of protein function
4. DNA _____ determines transgene _______
5. New _____ implanted into female egg

Answer 6

1. DNA
2. DNA; desired
3. gain ; loss
4. promoter ; expression
5. zygotes

Question 7

GFP, a neutral reporter, becomes fluorescent to study gene _____

Answer 7

expression

Question 8

Purkinje cell-specific promoters, functional proteins, cause ______ expressions in heavily populated Purkinje areas. Therefore, this leads to their ______.

Answer 8

inhibitor ; ablation

Question 9

The five steps for a knockout procedure:

1. target _____
2. introduce the ____ to _____ cells
3. use gRNA (_______ _______)
4. implant _____ ______ cells into embryo
5. _____ KO organism

Answer 9

1. gene
2. gene ; ES
3. homologous recombination
4. modified ES
5. breed chimera (animal with KO gene) twice

Question 10

Modified ES cells develop chimera and homogeneous KO animals due to cell ______, creating _____ and ____ ______ cells.

Answer 10

division ; equi and pluripotential

Question 11

List the 4 steps in KO embryonic stem cells:

1. ________ embryo inner cells
2. let cells _____ rapidly (like benign tumor)
3. mix with normal embryo to generate _________ cells
4. let cells ________ genomes

Answer 11

1. culture embryo inner cells
2. let cells grow rapidly (like benign tumor)
3. mix with normal embryo to generate pluripotential cells
4. let cells manipulate genomes

Question 12

Name the three types of KOs:

Answer 12

1. Global
2. Conditional
3. Inducible

Question 13

Global KO’s allow for spatial control in knockouts. This happens (early/late/anytime) in development.

Answer 13

early

knocks out gene in ALL tissue.

Question 14

Conditional KO’s allow for spatial and time control, choosing _____ in Knockouts. This happens (early/late/anytime) in development.

Answer 14

where ; when ; anytime

KO only in desired tissue

Question 15

Inducible KO’s allow for spatial and time control, choosing _____ and _____ in Knockouts. This happens (early/late/anytime) in development.

Answer 15

when ; where ; anytime

Question 16

Name the three steps in conditional KO:
1. _______ expresses CreER
2. _______ knocked-IN
3. genomes (KI/KO) specific tissue

Answer 16

1. promoter expresses CreER
2. loxP knocked-IN
3. genomes KO specific tissue

Question 17

T/F:

All cells in the body have a knocked-in loxP gene and Cre promoter.

Answer 17

TRUE

Question 18

In conditional gene targeting, loxP _____ the desired gene, allowing Cre to ____ it, causing a knock-out.

Answer 18

trap ; delete

Question 19

The four steps in inducible KO are as follows:

1. ____ binds to estrogen
2. ______ is injected
3. _____ moves in
4. ________ adds a new temporary function

Answer 19

CreER ; Tamoxifen ; CreER ; investigator

Question 20

the CRISPR/Cas9 system is like Cre/LoxP using ______ to destroy and invade virus’.

Answer 20

enzymes

Question 21

CRISPR/Cas9 takes a viral (section/strand) of (DNA/RNA) to make complementary (m/g)RNA. If the virus comes back, bacteria makes (r/g)RNA, Cas9 becomes a (reporter/hybrid) and cleaves DNA, inactivating the virus.

Answer 21

section ; DNA ; gRNA ; gRNA ; hyrbrid

Question 22

CRISPR/Cas9 is used for _____ editing with _______/_______

Answer 22

gene ; insertions/deletions

Question 23

Homology directed repair (HDR) corrects human _______

Answer 23

mutations

Crispr/Cas9 uses donor DNA (complements gRNA) to insert large DNA fragment into “cut” section.

Question 24

Nonhomologous end-joining (NHEJ) is an ____ prone way to _____ cells.

Answer 24

error ; repair

(generates small deletions/insertions on DNA ENDS)

Question 25

Two CRISPR goals are ___genesis and ____/____

Answer 25

1. mutagenesis (NHEJ) 2. Insertion/editing (HDR)

Question 26

T/F: CRISPR/Cas9 gene editing has the potential to correct human diseases. Provide a reason why.

Answer 26

TRUE. It can edit/mutate DNA in ES cells.

Question 27

RNA interference (RNAi) is a way to ______ RNA and reduce ____

Answer 27

degrade ; proteins

Question 28

For RNAi, you need to use knock-(downs/in).

Answer 28

knock-downs Targets and destroys RNA fragments

Question 29

The three steps in the RNAi process: 1. __RNA or __RNA complements knock___ RNA 2. __RNA makes __RNA, binds to RISC complex, becomes single stranded and complements __RNA 3. RISC and Dicer encode to be control mechanisms, ______ing and ______ing proteins

Answer 29

1. shRNA ; dsRNA ; down 2. shRNA ; siRNA, ; mRNA 3. regulating ; degrading

Question 30

The main point of RNAi is to make a __________ double strand RNA (___RNA) and knock-_____ a (desired/undesired) region

Answer 30

complementary ; (dsRNA) ; down ; undesired

Question 31

RNAi research is mostly used for _______ disease

Answer 31

Huntington's

Question 32

Antisense oligonucleotides (ASO) are short synthetic (DNA/mRNA) strands which (copy/insert) short (mRNA/DNA) sequences to make a ____-____ hybrid.

Answer 32

DNA ; copy ; mRNA ; RNA-DNA

Question 33

mRNA is called the ____ so naturally, ASOs are called the _____

Answer 33

sense ; antisense

Question 34

Oligo represents many __________ linked in a sequence

Answer 34

nucleotide