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Table 3 In vitro development of embryos injected with Cas9 mRNA and gRNAs knocking out simultaneously 5 genes

From: Generation of multi-gene knockout rabbits using the Cas9/gRNA system

Gene

gRNA/cas9 mRNA (ng/μL)

No. of injected zygotes

No. of blastocysts (%)

No. of sequenced blastocysts

No. of modified for a single gene (%)

No. of biallelic- modified (%)

No. of 2 or more genes- modified (%)

No. of 3 or

more genes- modified (%)

No. of 4 or more genes- modified (%)

No. of 5 genes- modified (%)

No. of 5 genes biallelic-modified (%)

IL2rg

   

12

11 (91.7)

11 (91.7) *

     

RAG1

   

15

14 (93.3)

12 (80.0)

     

RAG2

20/20/20/20/20/200

39

20 (51.3)

14

12 (85.7)

10 (71.4)

11/12 (91.7)

7/12 (58.3)

6/12 (50.0)

4/12 (33.3)

1/12 (8.33)*

TIKI1

   

14

9 (64.3)

6 (42.9)

     

ALB

   

19

11 (57.9)

5 (26.3)

     
  1. *indicates embryos without detecting WT sequence.