Table 1 Knock-in efficiency with various combinations.

From: CRISPR/Cas9-mediated reporter knock-in in mouse haploid embryonic stem cells

Combination No.

Cas9 type

sgRNA

HA of donor vector

The Venus-positive ratios

  

target

targeting seq. (nt)

 

Haploid (%)

Diploid (%)

1

Cas9

Actb

R(20)

Actb

18.0±1.2

8.2±0.6

2

Cas9

Actb

R(17)

Actb

21.2±0.9

10.3±0.3

3

Cas9n

Actb

L(20)+R(20)

Actb

30.6±1.5

14.3±0.4

4

Cas9n

Actb

L(20)+R(17)

Actb

31.1±1.3

14.4±0.6

5

Cas9n

Actb

R(20)

Actb

11.4±1.4

8.8±0.4

6

Cas9n

Actb

R(17)

Actb

4.0±0.5

3.3±0.2

7

Cas9n

Sox1

L(20)+R(17)

Actb

0.0±0.0

0.1±0.1

8

Cas9n

Actb

L(20)+R(17)

Sox1

0.1±0.1

0.0±0.0

9

No

  

Actb

0.0±0.0

0.0±0.0

 

The ratio of knock-in ESC lines

Haploid (%)

10

Cas9n

Sox1

L(20)+R(17)

Sox1

46.2 (n=26)

 
  1. The Venus-positive ratios were acquired using FACS 9 days after transfection and shown as the means ± SEM (n = 4). The knock-in efficiency at the Sox1 locus was calculated based on PCR analysis. A dot-plot graph and statistical analysis are shown in Fig. S5.