Homologous Recombination



  細胞類型 時間 成功率 脫靶率
Homologous recombination <5% >5K bp 約12個月 <25%
CRISPR/Cas >50% <5K bp 約6個月 >90%


  Process Contents Period
1 Construction of targeting vector 1. Understand customer demand of target genes to design targeting strategies 2. Cloning sequences of homologous arm from genome 3. Construction of targeting vector by homologous arm, positive and negative drug resistant cassettes 4. Linearization of targeting vector for electroporation Approx. 2 months
2 Establishment of homologous recombinant ES cell clones 5. Electroporation of targeting vector into ES cells. 6. Selection of drug-resistant ES cell colonies. 7. Screening the clones for homologous recombination-based deletion. 8. Expansion and establishment of ES cell lines with biallelic deletion. Approx. 2 months
3 Generation of chimeric mice 9. Blastocyst injection of modified ES cells 10.Embryo transfer to pseudopregnant female mouse 11.Generation of chimeric mice with max. 3 littermates Approx. 3 months
4 Generation of heterozygous mice 12.Generation of F1 heterozygous mice by germline transmission 13. Identification of F1 heterozygous mice by genotyping Approx. 3 months
5 Delivery 14.Transportation of mice Approx. 1 months


  • Knock out
  • Knock in
  • Over expression
  • Conditional mutant (Cre/Lox base)
  • Inducible system (Tet-On/ off, etc.)
  • Transgene


  • 每月進度報告
  • 模板載體定序報告
  • 細胞株定序報告
  • 基因型鑑定實驗步驟