Infection and Transfection of Human Embryonic Stem Cells

Infection and Transfection of Human Embryonic Stem Cells

Table S1 (Supplementary information).

List of available genetically modified hESC lines

Infection and transfection of human embryonic stem cells

hESC line / Transgene / Technique / Ref.
SNUhES3 / GFP / Transient transfection using Fugene 6 / 1
H1 / GFP / Transient transfection using nucleofection (Amaxa) / 2
H9 / GFP / Transient transfection with Fugene 6 / 3
H1 / GFP / Stable transfection with Fugene 6
(neomycine selection) / 4
H9 / GFP
dsRED2 / Stable transfection with Lipofectamine 2000, Exgen 500
(puromycin selection) / 5
SNUhES3
(D) /

Noggin

/ Stable transfection using Fugene 6
(G418 selection) / 1
H9 / Nodal / Stable transfection with Lipofectamin 2000
(puromycine selection) / 6
H9 / Lefty2
Cerberus-Short
(Constitutive inhibitors of Nodal) / Stable transfection with Lipofectamin 2000
(puromycine selection) / 7
H9 / HSV-TK / Stable transfection with Exgen 500
(neomycin selection) / 8
H1, H9 / EBNA1-neoR
GFP
/ Stable transfection of EBNA1 cDNA with Fugene 6 (neomycin selection)
Episomal EBV-based plasmid expression of GFP / 9
H9 / Pdx1
FoxA2 / Stable transfection with Exgen 500
(Neomycin selection) / 10
H9.2 / hcRed (flanked with loxP sites)
GFP / Stable transfection by electroporation
(neomycin selection) / 11
H7 /
GFP Neo / MSCV Retrovirus / 3
H1, H7, H9
(D) / hTERT / pBABE-MLV Retrovirus / 12
H1, H9
(D) / HOXB4 / MIEV Retrovirus / 13
H9
(D) / HPV16E6E7
hTERT / Retrovirus / 14
H9 / -gal / Adenovirus and adeno-associated virus :
Ad5, AAV2, AAV4 and AAV5 / 15
H1 / GFP / Lentivirus / 16
HES-1 / GFP / Lentivirus / 17
H9 / GFP / Lentivirus / 18
hES-1 / GFP-IRES-PuroR / Lentivirus
(comparison of several IRES and promoters) / 19
H1 / GFP / Lentivirus
(Comparison of different pseudotypes) / 20
H1 / GFP / Lentivirus / 21
(D) indicates DNA transfection to differentiated hESC
Knock-down
hES line / Downregulated gene / Method / Ref.
H9, H1 / RNAi against Oct4 / Transient transfection with Lipofectamin 2000 / 22
H9 / RNAi against GFP
RNAi against dsRED / Transient transfection with Oligofectamine / 5
H1,
hES-NCL1 / RNAi against Nanog / Transient transfection with Lipofectamin 2000 / 23
H1 / RNAi against Oct4 / Transient transfection with Oligofectamin / 24
H7, H14 / RNAi against Oct4
RNAi against
2-microglobulin / Stable transfection with Exgen 500
(puromycine selection) / 25
H9 / RNAi against HPRT / Stable transfection with Oligofectamin, Exgen 500
(Puromycine selection) / 5
H1 / RNAi against GFP / Lentivirus / 26
H9 / RNAi against Oct4
RNAi against Nanog / Lentivirus / 27
Knockout (Homologous recombination)
hES line / Gene / Method / Ref.

H13

/

Knockout of HPRT1

/ Transfection with Exgen 500
(Hygromycin selection) /

28

H1.1

/

Knockout of HPRT1

/ Transfection with electroporation
(Neomycin selection) /

29

Reporter cell lines

hES line / Downregulated gene / Method / Ref.

H1.1

/

Oct-4 knock-in reporterOct4+/IRES-GFP-IRES-NeoR

/

Transfection with electroporation(Homologous recombination, selection with Neomycin)

/

29

H9 / Rex1 reporter
Rex1::GFP / Stable transfection with Exgen 500
(Neomycin selection) / 30
H9 / Albumin reporter
ALB::GFP / Stable transfection with Exgen 500
(Neomycin selection) / 31
H9 / Rat Insulin1
and mouse Pdx1 reporter
Ins::GFP
Pdx1::GFP / Stable transfection with Exgen 500
(Neomycin selection) / 10
H1 / Oct4 reporter
Oct4::GFP / Stable transfection with Fugene 6
(Neomycin selection) / 24

References for Table S1

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