CRISPR Gene Editing in Stem Cells

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0
10
20
30
40
50
60
0
10
20
30
40
50
60
70
0
1
2
3
4
5
6
7
0
0.5
1
1.5
2
2.5
3
noneforsknoneforsknoneforsknoneforsk
0
0.5
1
1.5
2
2.5
3
3.5
0
0.5
1
1.5
2
2.5
LGHGLG HGLGHGLG HG
0
10
20
30
40
50
60
70
80
90
0.01 0.1 1 10 100
0
20
40
60
80
100
120
140
160
180
200
0.1 1 10 100 1000 % of PI
+ INS
+ in INS
+ cells
# INS
+cells
Concentration (uM)
A B C
E
22.6
23.4
43.1
39.1
21.2
21.3
62.5
43.4
Annexin V
T5224 DMSO
7AAD
ctrl-g glu ctrl-g glu
% of Annexin V
+cells
in insulin-GFP
+ cells
DMSO T5224
n.s. G
D
T5224 DMSO
wt CDKAL1-/-
ctrl-p palm ctrl-p palm
Fold Change Fold Change
***
***
*
n.s.
**
**
*** ****
*
n.s.
***
n.s.
DMSO T5224 DMSO T5224 DMSO T5224 DMSO T5224
wt CDKAL1-/-
DMSO T5224 wt CDKAL1-/-
DMSO T5224
H I
PI
insulin
DAPI
n.s.
**
n.s.
wt CDKAL1 -/-
wt CDKAL1-/-
wt CDKAL1-/-
*
% of PI
+ INS
+ in INS
+ cells
Human c-peptide (% of content) Human c-peptide (% of content)
16.1
16.2
39.6
40.8
17.1
17.4
58.4
42.5
wt CDKAL1-/-
T5224 DMSO
T5224 DMSO
wt CDKAL1-/-
ctrl-g glu ctrl-g glu
****
n.s.
n.s.
n.s.
n.s.
***
n.s.
n.s.
n.s.
ctrl-p palm ctrl-p palm
F
ctrl-g glu ctrl-g glu
wt CDKAL1 -/-
DMSO T5224
n.s.
****
n.s. n.s.
n.s.
n.s.
***
n.s.
n.s.
n.s.
ctrl-p palm ctrl-p palm
ctrl-p palm ctrl-p palm
ctrl-g glu ctrl-g glu
wt CDKAL1 -/- wt CDKAL1-/- wt CDKAL1-/-
(legend on next page)
Cell Stem Cell 19, 326–340, September 1, 2016 333