迅速高效的基因編輯技術,利用gRNA辨識基因編輯位置,引導Cas9至該位置後造成DNA雙股斷裂, 以進行後續的基因編輯。
該技術可以除了單純的DNA雙股斷裂造成indel的發生,亦可以利用HDR template來達成想要的突變形式,如:基因敲除、基因敲入、點突變、基因轉殖。
脫靶率 | 可編輯範圍 | 時間 | 成功率 | |
CRISPR/Cas | >50% | <5K bp | 約6個月 | >90% |
Homologous recombination | <5% | >5K bp | 約12個月 | <25% |
Process | Contents | Period | |
1 | Construction of targeting vector | 1. Understand customer demand of target genes to select targeting sites 2. Cloning targeting sequences for guide RNA 3. Synthesis of guide RNA and cas9 mRNA |
Approx. 1 months |
---|---|---|---|
2 | Generation of F0 mice | 4.Injection of guide RNA and cas9 mRNA into zygotes 5.Embryo transfer to pseudopregnant female mouse 6.Generation of F0 mice with max. 3 littermates |
Approx. 2 months |
3 | Generation of F1 heterozygous mice | 7.Generation of F1 heterozygous mice by germline transmission 8.Identification of F1 heterozygous mice by genotyping |
Approx. 3 months |
4 | Delivery | 9.Transportation of mice | Approx. 1 month |