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Table 3 Generation of a GckrP446L point mutation

From: Application of long single-stranded DNA donors in genome editing: generation and validation of mouse mutants

      

F0 with:

MS

Donor type

Guide ID(s)

Donor ID

Embryos transferred

F0 biopsied (birth rate)

Mutation

Correct mutation

SM only

SM only and rearranged

NHEJ alleles

Random integration

1

ssODN

20

Gckrdonor_20

80

12 (15%)

0

0

0

0

0

n.d.

2

ssODN

3

Gckrdonor_2

80

21 (26%)

12

0

2

2

9

n.d.

3

ssODN

3

Gckrdonor_2

70

13 (19%)

4

0

0

0

4

n.d.

4

ssODN

3

Gckrdonor_2

135

18 (13%)

9

0

2

1

7

n.d.

5

ssODN

3

Gckrdonor_2

42

10 (23%)

3

0

0

0

3

n.d.

6

ssODN

3

Gckrdonor_3

121

8 (7%)

5

0

1

1

3

n.d.

7

ssODN

3

Gckrdonor_3

112

8 (7%)

3

0

0

0

3

n.d.

1

lssDNA

5.2, 3.1

Gckr_P446L_lss

210

22 (10%)

14

8

1

2

7

0/2

  1. The table shows the numbers of embryos and animals involved in mutagenesis attempts employing the injection of CRISPR/Cas9 reagents and oligonucleotides or lssDNA donors. The percentage of transferred embryos yielding live animals at weaning is shown in parentheses. The outcome of these attempts is also summarized. Note that sgRNA_20 was employed for the first microinjection session with ssODN_20 and substituted to sgRNA_3 and relevant donor ssODNs for subsequent sessions, as it was confirmed to be inactive. Sequencing data from this project are displayed in Fig. 4 (additional raw sequencing data are provided in Additional file 16)
  2. MS microinjection session, n.d. not determined SM silent mutation