Tet Inducible Gene Expression Stable Cell Line
VectorBuilder는 background 발현을 최소화하고 GOI를 높은 유도발현으로 Tet inducible 유전자 발현 안정 세포주를 맞춤형으로 제작할 수 있습니다. 또한 Tet regulatory 단백질(예: rtTA, tTS/rtTA, tet3G 등)을 발현하는 안정적인 세포주를 디자인할 수 있으며, 유연한 실험 설계와 신뢰할 수 있는 Tet inducible 유전자 발현을 위해 TRE에 의해 구동되는 GOI를 운반하는 바이러스/플라스미드로 transduction/transfection 될 수 있습니다. Tet-On 안정 세포주의 경우 GOI 유도는 RT-qPCR에 의해 검증됩니다. 또한, 최종 세포주 제품을 출시하기 전에 무균 테스트, mycoplasma 검출 등 일련의 표준 QC 분석을 수행합니다.
중점 사항
- Switch-like gene activation: VectorBuilder의 벡터 시스템은 유전자 발현을 제어하고 background 발현을 최소화하며 높은 감도를 나타내기 위해 tetracycline-regulated on-and-off switch를 허용합니다.
- High-level expression: TRE 프로모터는 유도된 상태에서 매우 높은 수준의 유전자 발현을 유도할 수 있습니다.
- Rapid turnaround: 벡터 디자인부터 시작하여 빠르면 9주 만에 제작할 수 있습니다.
서비스 상세
Figure 1. Typical workflow of Tet inducible gene expression stable cell line production.
가격 및 소요시간
| Service Type | Deliverable | Price (USD)* | Turnaround |
|---|---|---|---|
| Tet-inducible gene expression | Mixed pool (>106 cells/vial, 2 vials) | From $3,999 | 9-15 weeks |
| Two single clones (>106 cells/vial, 2 vials per clone) | From $4,999 | 14-20 weeks |
* Additional charge will apply for extra single clones or vials.
QC assays
| Assays | Methods |
|---|---|
| Tet-induced overexpression validation (default) | RT-qPCR |
| Expression test (add-on) | WB, IF, FACS |
| Sterility (default) | PCR for mycoplasma detection, bioburden test for sterility detection |
다운스트림 서비스
VectorBuilder는 proliferation, apoptosis, migration, viability, cytotoxicity 등에 대한 분석을 포함하여 제작된 세포주의 다양한 phenotype 평가 및 기능 검증을 수행할 수 있습니다.
사례 연구
Figure 2. Tet-inducible EGFP expression in 293T cells. The tTS/rtTA stable expression 293T cells were transduced with lentivirus carrying TRE-driven EGFP at an MOI of 1. EGFP expression was measured in the absence (Dox-) or in the presence (Dox+) of doxycycline (tetracycline derivative, 10 ng/ml) using (A) microscopy, (B) flow cytometry, and (C) western blot at 24 h and 48 h post-transduction. MFI, median fluorescence intensity.
How to Order
FAQ
The tetracycline inducible gene expression system is a powerful tool to control the timing of gene expression in mammalian cells. Our Tet-On vector systems are designed to achieve nearly complete silencing of a GOI in the absence of tetracycline, and strong, rapid expression in response to the addition of tetracycline. This is achieved through a multicomponent system which incorporates active silencing by the tTS protein in the absence of tetracycline and strong activation by the rtTA protein in the presence of tetracycline. In the absence of tetracycline, the tTS protein derived from the fusion of TetR (Tet repressor protein) and KRAB-AB (the transcriptional repressor domain of Kid-1 protein) binds to the TRE promoter, leading to the active suppression of gene transcription. The rtTA protein, on the other hand, derived from the fusion of a rTetR (a mutant Tet repressor) and VP16 (the transcription activator domain of virion protein 16 of herpes simplex virus), binds to the TRE promoter to activate gene transcription only in the presence of tetracycline. Our Tet-Off vector system can be used for inhibiting target gene expression in the presence of tetracycline regulated by the binding of the tTA protein to the TRE promoter.