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) |