Homologous recombination

同源重組技術

來自諾貝爾獎實驗室的精準編輯技術,利用同源重組技術可以將指定的區域置換成想編輯的形式,擅長進行無脫靶副作用的大片段基因編輯。


技術優勢

  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%

HR基因編輯鼠-製作流程

  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

客製化服務

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

我們將提供您

  • 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

常見問題

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案例分享

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