部位特異性遺伝子修飾は普段内因性DNAと外因性DNAの相同組換え(HR)を行っています。ゲノム編集技術はマウスの胚性幹細胞(ESC)中のDNA相同組換え技術(HR)に由来し、DNAヌクレアーゼゲノム編集技術(ジンクフィンガーヌクレアーゼ(ZFN) と新しいCRISPR-Cas9技術の出現に伴って、新たな段階へと発展しました。ZFNとTALENを比べて見ると、CRISPR-Cas9技術はより効率的で、作製時間を短縮し、生産コストを低減するだけでなく、種間の制約を打ち破り、患者組織内の遺伝子を直接編集の潜在力もあります。

バイオサイトジェンはCRISPR-Cas9を基盤とするEGE™システムを開発しました。このシステムはラット/マウス及び細胞株のDNA断片(5kb以上)のノックイン効率を20倍近く向上させています。EGE™技術で98%のプロジェクトを成功し、ゲノム編集動物及び細胞株モデルの作製期間を非常に短縮しています。

TechnologyAdvantagesApplicationsTimeline
ESC/HR-based Gene EditingCRISPR / EGE™High efficiency; large gene knockout/knockin capabilities; simultaneously targets multiple genes; applicable to many species; easy to construct. Off-target effects can be reduced by choosing the appropriate sgRNA and eliminated by breeding; Southern blot to screen out random insertions.Conventional KO mice/rats6-8 months
Conditional KO mice/rats
ROSA26 locus gene KI mice/rats
Gene KI/ Mutation mice/rats
Humanized mice/rats
Reporter/KO-then-cKO mice/rats
Normal cell lines KO/KI
hESC/iPS cells KO/KI
ESC/HR-based Gene EditingAccurate, precise and reliably established, by far the only targeting technology which can meet the requirements of almost all genome modifications; but ES cells are required (only mouse cells are efficient for this technique), so it cannot be applied to other animal models); time consuming; heavy workload; high costConventional knockout (KO) mice7-11 months
Conditional KO mice
ROSA26 locus gene knockin (KI) mice
Gene KI/ Mutation mice
Humanized mice
Reporter/KO-then-cKO mice
Transgenic TechnologyTraditional transgenic technology is welcomed because the fast timeline low cost, super large fragment insertion, etc. But as the advance of new gene targeting technologies, especially EGE™, above advantages are not very obvious anymore. Especially random insertion, instability, large number founders can cost more time and money sometimesTransgenic mice/rats2-3 months for F0
Chromosome EngineeringGene cluster replacement at the scale of megabase. A unique technology for specific applications such as antibody gene cluster humanization.MouseUpon request
Others ServicesBesides the full-scale gene editing services, Biocytogen also provide milestone services relating with gene editing.CRISPR plasmid construction; CRISPR activity assay; Donor plasmid constructionUpon request

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