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Table 1 Summary of i-GONAD using knock-in enhancers

From: Modification of improved-genome editing via oviductal nucleic acids delivery (i-GONAD)-mediated knock-in in rats

crRNA used

Enhancers1

No. of pregnant rats/no. of treated rats

Total number of fetuses obtained

No. of fetuses or pups carrying pigmented eyes (successful KI [%])

No. of fetuses showing indels (%)

No. of fetuses showing mosaic mutations (%)

No. of fetuses showing no mutation (%)

1

–

4/7

40

2 (5)

16 (40)

5 (13)

23 (58)

 

1 µM SCR7

6/6

27

2 (7)

11 (41)

6 (22)

16 (59)

 

10 µM SCR7

4/4

23

2 (9)

10 (43)

5 (22)

13 (57)

 

100 µM SCR7

3/3

32

2 (6)

16 (50)

4 (13)

15 (47)

 

4 µM EP Enhancer

2/4

12

0 (0)

4 (33)

1 (8)

8 (67)

 

30 µM AZT

4/4

21

0 (0)

10 (48)

1 (5)

11 (52)

 

5 µM L755

4/4

26

3 (12)

9 (35)

4 (15)

17 (65)

 

15 µM L755

5/6

33

4 (12)

14 (42)

2 (6)

17 (52)

 

15 µM RS-1

6/9

25

2 (8)

12 (48)

2 (8)

13 (52)

 

30 µM HDR Enhancer

4/4

28

3 (11)

16 (57)

8 (29)

12 (43)

 

Mixture of four enhancers2

3/3

21

2 (10)

11 (52)

3 (14)

9 (43)

2

–

4/4

21

5 (24)

18 (86)

2 (10)

3 (14)

 

10 µM SCR7

3/3

30

5 (17)

16 (53)

4 (13)

12 (40)

 

5 µM L755

3/4

18

3 (17)

9 (50)

2 (11)

8 (44)

 

15 µM L755

3/3

20

6 (30)

9 (45)

3 (15)

6 (30)

 

30 µM HDR Enhancer

4/4

22

3 (14)2

10 (45)

2 (9)

11 (50)

3

–

5/5

38

0 (0)

7 (18)

0 (0)

31 (82)

1 + 2 + 3

–

8/9

62

7 (11)

25 (40)

9 (15)

35 (56)

  1. 1EP Enhancer, Alt-R® Cas9 Electroporation Enhancer; AZT, azidothymidine; L755, L755,507; RS-1, RAD51-stimulatory compound 1; HDR Enhancer, Alt-R® HDR Enhancer
  2. 2Mixture of 10 µM SCR7, 5 µM L755, 15 µM RS-1, and 30 µM HDR Enhancer
  3. –in vivo EP in the absence of any reagent