Evaluation of DNA damage, cellular toxicity and inflammatory markers following saCas9 and gRNAs delivery by AAV2 in kidney organoids. |
AAV2/2-SaCas9 + AAV2/2-gRNA |
Compartmental DNA damage was calculated as the percent of YH2AX co-staining among LTL, CHD1 and MEIS1/2/3 positive cells |
|
Biomarker Detection |
15000003443 |
|
SaCas9 (AAV-SaCas9) |
AAV |
Viral vector |
DMD low gRNA-2 |
AAV2-gRNA; |
|
Human |
Human kidney organoid (stem cell-derived) |
|
virus |
|
MOI=10^5 |
|
|
|
7 days |
AAV vector encoded |
|
|
|
RRID:AB_2750570 |
Active Motif Cat# 39795 |
Mouse anti-meis1/2/3 |
|
RRID:AB_2118010 |
Cell Signaling Technology Cat# 2577 |
Rabbit anti-Phospho-Histone H2A.X (Ser139) |
|
RRID:AB_307284 |
Abcam Cat# ab9498 |
Mouse anti-CD31 antibody |
|
RRID:AB_2336558 |
Vector Laboratories Cat# B-1325 |
Lotus tetragonolobus lectin (LTL) |
|
RRID:AB_2116561 |
R and D Systems Cat# AF1750 |
Goat anti-KIM1/HAVCR |
|
Measured Values |
Samples Size: | 3 |
|
Mean |
69.29  |
1 |
83.33333  |
2 |
66.66667  |
3 |
50  |
4 |
75  |
5 |
75  |
6 |
75  |
7 |
60  |
Mean |
55.07  |
1 |
43.3333333333333  |
2 |
77.4193548387097  |
3 |
44.4444444444444  |
Mean |
89.34  |
1 |
87.1508379888268  |
2 |
94.2028985507246  |
3 |
86.6666666666667  |
Mean |
34.87  |
1 |
29.375  |
2 |
33.6538461538462  |
3 |
41.5770609318996  |
Mean |
487.92  |
1 |
495.417375758954  |
2 |
464.933192143006  |
3 |
526.17876439772  |
4 |
476.437321338346  |
5 |
497.545747150637  |
6 |
453.317100769354  |
7 |
522.919777936497  |
8 |
466.579803799361  |
Mean |
6.2  |
1 |
7.13986096462846  |
2 |
7.51252056707063  |
3 |
3.84216984250873  |
4 |
5.13790801017772  |
5 |
6.91414028649866  |
6 |
5.20233895635352  |
7 |
6.79682820460547  |
8 |
7.06756386236927  |
Mean |
3.05  |
1 |
3.04035320059101  |
2 |
3.45685924657181  |
3 |
2.50921630343338  |
4 |
2.51575347278385  |
5 |
3.02762559397732  |
6 |
2.63136714995803  |
7 |
3.56041744685856  |
8 |
3.69759523331727  |
Mean |
101.69  |
1 |
73.0450520275434  |
2 |
247.799688076254  |
3 |
57.3500937297986  |
4 |
51.5853862679727  |
5 |
104.236737882331  |
6 |
50.1516171305266  |
7 |
102.742313385041  |
8 |
126.625898599655  |
Mean |
10.69  |
1 |
10.552638994931  |
2 |
10.0996567940629  |
3 |
6.94973325618479  |
4 |
6.54733914189619  |
5 |
16.9785383180146  |
6 |
6.82945998412296  |
7 |
17.9601172079959  |
8 |
9.57669773080734  |
Mean |
3682.05  |
1 |
2240.35305387958  |
2 |
3082.66840339688  |
3 |
2182.40694910355  |
4 |
2029.08363507414  |
5 |
5895.0685121659  |
6 |
2129.72429900216  |
7 |
6258.05943091146  |
8 |
5639.0439532151  |