GV10: Stem Cells & Gene Manipulation

Question 1

what are stem cells

Answer 1

unspecialised cells that can proliferate and differentiate into many other cell types

Question 2

what are the three major types of stem cells

Answer 2

adult, embryonic, induced pluripotent

Question 3

what are the transcription factors used in regulation of mouse ES cell self-renewal

Answer 3

oct-4, nanog, sox-2

Question 4

what are the characteristics of induced pluripotent stem cells

Answer 4

no ethical issue since derived from adult cells, can proliferate indefinitely, pluripotent, chances of graft-versus-host rejection is reduced since these are derived from individuals to be treated, may develop into cancers

Question 5

what are the advantages of embryonic stem cells

Answer 5

renewable, pluripotent, allogenic

Question 6

what are the disadvantages of embryonic stem cells

Answer 6

ethical concerns

Question 7

what are the advantages of adult stem cells

Answer 7

multipotent, allogenic or autologous, no ethical concerns

Question 8

what are the disadvantages of adult stem cells

Answer 8

rare, limited supply

Question 9

what are the advantages of iPS

Answer 9

renewable, pluripotent, autologous, no ethical concerns

Question 10

which species can be used as animal models

Answer 10

fruit flies, worms, fish, amphibians, rodents, primates, farm animals

Question 11

what are transgenic mice

Answer 11

contain additional, artificially-introduced genetic material in every cell
genetic material added to genome so animal makes excess amounts of the gene/protein encoded by this material

Question 12

why are mice the animal of choice

Answer 12

lower cost, small size, rapid reproduction rates, sequence of the entire genome known

Question 13

how are transgenic mice produced

Answer 13

embryonic stem cells are transfected with virus, plasmid or vector containing gene of interest (random insertion)
select ES cell expressing transgene by gRT-PCR

Question 14

what is gene targeting (gene knockout)

Answer 14

introduction of a defined mutation into a specific gene to study gene function
achieved by homologous recombination

Question 15

what are the limitations of gene targeting

Answer 15

requires detailed understanding of the intron-exon structure of target gene
homologous recombination in mammalian cells is rare
function of gene during development may be different from its function in the adult