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Table 1 Generation of conditional knock-out mice using lssDNA

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

 

Number of F0s (with)

Number of lines with:

Gene name

MSa

Biopsied

Mutation

cKO alleles

Illegitimate repair

Exon deletion

Indels

Progenyb

GLT cKO

GLT exon deletion

GLT illegitimate repair

Additional insertion(s) detected/tested by ddPCR

QC pass

Syt7

1

17

10

2

1

3c

5

2 (+ 2)

2

0 (+ 2)

1 (+ 0)

1 (+ 1) / 2 (+ 2)

2 (+ 2)f

Ikzf2

1

17

5

1

1

3

5

1 (+ 1)

1

1 (+ 0)

1 (+ 1)

0 (+ 0) / 1 (+ 1)

1 (+ 1)

 

2

6

4

0

1

3

4

0 (+ 2)

–

– (+ 2)

– (+ 0)

– (+ 2) / – (+ 2)

– (+ 1)f

Syt4

1

26

16

0

10

1

11

0 (+ 1)

–

– (+ 1)

– (+ 1)

– (+ 0) / – (+ 1)

– (+ 1)

 

2

40

16

1

9

1

9

2 (+ 1)

1

0 (+ 0)

2 (+ 1)

0 (+ 0) / 1 (+ 0)

1 (+ 0)

Usp45

1

19

7

1

0

2

6

1 (+ 1)

0

0 (+ 0)

1 (+ 0)

–

0 (+ 0)

 

2

6

1

0

1

0

1

0

–

–

–

–

–

Rapgef5

1

30

13

1d

5

6

8

0 (+ 2)

–d

– (+ 1)

– (+ 0)

0 (+ 0) / 0 (+ 1)

– (+ 1)

Cx3cl1

1

3

3

0

3

1

3

0

–

–

–

–

–

 

2

16

8

3

3

6

6

3

1

0

1

0 (+ 0) / 1 (+ 0)

1 (+ 0)g

6430573F11Rik

1

9

5

0

2

3

3

0 (+ 1)

–

– (+ 1)

–

– (+ 0) / – (+ 1)

– (+ 1)

 

2

21

14

2e

1

7

6

2e

0

–

2

–

–

 

3

28

3

0

1

3

2

0

–

–

–

–

–

 

4

5

3

0

1

1

3

0

–

–

–

–

–

Acvr2b

1

11

8

0

1

0

7

0

–

–

–

–

–

Inpp5k

1

4

2

0

1

0

1

0

–

–

–

–

–

 

2

21

11

2

2

6

8

2

0

0 (+ 0)

2

–

–

  1. The table summarizes the numbers of animals involved in the generation of conditional alleles employing CRISPR/Cas9 reagents and lssDNA donors and the outcome of mutagenesis. The table also shows the outcome of the breeding of positive founders, each generating a new line. Further summaries and sequencing data for each of the projects detailed in Table 1 are shown in Additional file 2: Figure S1, Additional file 3: Figure S2, Additional file 4: Figure S3, Additional file 5: Figure S4, Additional file 6: Figure S5, Additional file 7: Figure S6, Additional file 8: Figure S7, Additional file 9: Figure S8, Additional file 10: Figure S9 and Additional file 11: Figure S10
  2. aMS microinjection session
  3. bFounders with progeny for transmission of floxed allele (in brackets founders bred for transmission of deletion allele only)
  4. cOne additional deletion allele identified by sequencing
  5. dFounder with floxed allele died before mating
  6. eOne founder with floxed allele with point mutation in intron found in F0
  7. fRandom insertion of donor sequence detected by ddPCR in line but not transmitted to all F1 animals of interest, e.g. cKO/exon deletion
  8. gRandom insertion detected but not associated with allele of interest, e.g. cKO/exon deletion
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