Lecture 23

Question 1

Introducing genes to treat diseases

Answer 1

gene therapy

Question 2

Germ-line gene therapy: genes are introduced into the germ cells to – future generations.

Answer 2

genetically alter

Question 3

Somatic cell gene therapy: normal genes are added to the – of at least some of the body’s cells to ameliorate the effects of a defective gene

Answer 3

nuclei

Question 4

Some Goals of Gene Therapy:

Replace – genes with normal genes to cure diseases

Answer 4

defective

Question 5

Some Goals of Gene Therapy: introduce genes which will enhance –

Answer 5

immune defenses

Question 6

Some Goals of Gene Therapy: introduce – into cancer cells

Answer 6

suicide genes

Question 7

Some Goals of Gene Therapy: alter – of genes in the cell

Answer 7

regulation

Question 8

Some Goals of Gene Therapy: stop the replication of –

Answer 8

viruses

Question 9

Genes can be introduced to cells in culture (ex vivo) or –

Answer 9

in the patient (in vivo)

Question 10

+ Adenovirus

Answer 10

Infects many cell types

Question 11

• Adenovirus

Answer 11

Does not integrate into host genome and can be lost.

Question 12

+ Adeno-associated virus (AAV)

Answer 12

Integrates into host genome at specific site and cannot be lost

Question 13

• Adeno-associated virus (AAV)

Answer 13

difficult to work with

Question 14

• Simple retrovirus

Answer 14

integrate near genes

Question 15

+ complex retrovirus (lentivirus)

Answer 15

Integrates into host genome and cannot be lost

Integration is not random – occurs away from genes

Question 16

+ Herpes Simplex Virus (HSV)

Answer 16

DNA stays in nucleus without integrating into genome

Question 17

• Herpes Simplex Virus (HSV)

Answer 17

only infects nervous system cells

Question 18

Non-viral methods of gene transfer

Answer 18

liposome/lipolpex or naked DNA

Question 19

– are spheres of lipid molecules surrounding an aqueous interior (bilayer); limited amount of DNA will fit in it

Answer 19

liposome

Question 20

– are lipids with positive charge so attract both DNA and cell wall
No immune response, but not as efficient as virus at transferring DNA

Answer 20

lipolplex

Question 21

Direct injection of – into muscle cells was found to be expressed
Could be used to make insulin, blood clotting factors or as vaccines to viruses.

Answer 21

naked DNA

Question 22

Ex vivo usually targets – which are stem cells that generate all other blood cells

Answer 22

hematopoietic stem cells (HSC)

Question 23

X-linked severe combined immunodeficiency (SCID-X1)
β-thalessemia
WAS – an immune difficiency disorder
are associated with

Answer 23

HSC

Question 24

is an RNA-mediated mechanism for regulating gene expression in eukaryotes (regularly used to silence specific genes)

Answer 24

RNA interference

Question 25

RNAi is a cellular defense against viruses and --

Answer 25

transposable elements

Question 26

viral RNA uses -- to become dsRNA virus

Answer 26

RNA dependent RNA polymerase

Question 27

-- cuts dsRNA into 21-25 bp fragments

Answer 27

Dicer

Question 28

siRNA fragments created by Dicer bind to -- which denatures one of the two strands

Answer 28

RISC

Question 29

guide RNA remains bound to RISC and then the -- slices other RNA that is complementary to the guide strand

Answer 29

argonaute

Question 30

RNAi is used in research to test hypothesis of gene function this is known as --

Answer 30

knock-down

Question 31

RNAi is used in therapeutics to -- RNA of overexpressed genes

Answer 31

degrade

Question 32

RNA is used in therapeutics to target specific --

Answer 32

viruses (HIV)

Question 33

T/F: RNAi destroys genome

Answer 33

false

Question 34

RNAi methods: -- of siRNAs directly into cells

Answer 34

transfection

Question 35

RNAi method: introduction of -- synthesizes RNA identical to endogenous gene of interest (siRNAs made in organism)

Answer 35

vectors

Question 36

T/F: small interfering RNA (siRNA) come from exogenous sources or from other endogenous transcription

Answer 36

true

Question 37

Abnormal over-expression of -- lead to diseases such as age-related macular degeneration (AMD)

Answer 37

VEGF (vascular endothelial growth factor)

Question 38

VEGF is a protein required for blood vessel formation Excessive blood vessels damage retina -- VEGF should solve the problem

Answer 38

silencing

Question 39

problems associated with RNAi include the -- of sRNAs and adequate delivery systems

Answer 39

stability

Question 40

In RNAi silencing, specificity is provided by --

Answer 40

siRNA

Question 41

go in and make changes at a particular site, generally not replacing large amounts

Answer 41

genome editing

Question 42

ancient bacterial immune system which identifies the DNA of invading viruses and sends in an endonuclease, like Cas9, to shew it up

Answer 42

CRISPRS

Question 43

-- can be reprogrammed to recognize nearly any sequence of DNA just need the right RNA sequence to search a strand of DNA for where to cut

Answer 43

Cas9

Question 44

The -- regions are homologous to viruses

Answer 44

spacer

Question 45

The -- are important for binding the Cascade complex

Answer 45

repeats

Question 46

Spacer RNA directs complex to -- DNA or RNA

Answer 46

homologous

Question 47

In E.coli system, cascade complex has -- activity

Answer 47

Dnase

Question 48

In Pyrococcus furiosus system, Cmr-complex has -- activity

Answer 48

Rnase

Question 49

One can now use the CRISPR system in eukaryotes to do gene knockouts, gene replacements, and --

Answer 49

gene “drive”

Question 50

In crop plants, the genome can be edited and then the -- can be segregated away, leaving the plant transgene free

Answer 50

Cas9 plasmid

Question 51

selfish genes that appear more frequently in offspring than normal genes, which have about a 50-50 chance of being passed on

Answer 51

gene drives

Question 52

T/F: A gene drive always ends up in all offspring, even if only one parent has it; if given enough generations, it will eventually spread through the entire population

Answer 52

true

Question 53

Gene drive: Fight Malaria by changing the mosquitos that are part of their life cycle. One group modified females to become --

Answer 53

sterile

Question 54

Gene drive: Fight Malaria by changing the mosquitos that are part of their life cycle. The other group modified mosquitos by inserting two -- directed against the malarial parasite.

Answer 54

antibody genes

Question 55

Transgenic to fight Zika virus larvae died due to high expression of --

Answer 55

lethal transgene