Supplementary Figure 1; Large Transposon TnT
ITR CCTG300 ITR
KT2/HSA-CCTG300 9070 bpProbe
E E E
+ CpG
- CpG
200 bp
300 bp
B
C
511
510
509
508
507
506
505
504
503
502
501
L
Supplementary Figure 2; Rat genotype and transgene expression
ITR P BSD:GFP pA ITRmiR30-Rab38E
KT2/KDRab38 3534 bp
A
Supplementary Figure 3; F0 methylation
RegionSV
PSV
PSV
PSV
PSV
PSV
PSV
PSV
PSV
P
Founder # 2 4 5 6 8 13 20 74 79
# Inserts 4 4 4 2 4* 3 4 1 1
0
20
40
60
80
100
Per
cen
t C
pG
’s M
eth
ylat
ed
106
Supplementary Figure 4; LM-PCR founder 106
2.7 kb
4.8 kb
1 2 3 4 5 6 7 8 9 100
1
2
3
4
5
6
7
8
±CpG ; TO
KT2H-CD40Ig + CpG
KT2H-CD40Ig - CpG
sc KT2H-CD40Ig + CpG
sc KT2H-CD40Ig - CpG
T2/sh_mCFTR1 + CpG
T2/sh_mCFTR - CpG
KT2/HAS-CCTG300 + CpG
KT2/HAS-CCTG300 - CpG
1 2 3 4 5 6 70
1
2
3
4
5
6
7
8
±CpG; TMC
KT2H-CD40Ig + CpG
KT2H-CD40Ig -CpG
sc KT2H-CD40Ig + CpG
sc KT2H-CD40Ig - CpG
T2/P2_mCFTR1 + CpG
T2/sh_mCFTR - CpG
KT2/HAS-CCTG300 + CpG
KT2/HAS-CCTG300 - CpG
Supplementary Figure 5; Insert copy number distribution
# F
ound
ers
# F
ound
ers
Copy number
Copy number
B
A
Non-methylated
Non-methylated
Small Intestine Puro mRNA
Liver shCFTR1
Supplementary Figure 6; Broad transgene expression range in T2/shP2-mCFTR transgenic animals
B
A