- 최적화를 통한 고발현과 고역가 실현
- 단지 1μl의 순수한 바이러스 상층액 만으로도 10 cm dish의 모든 세포에 도입 가능
- WPRE과 cPPT element가 도입효율과 발현율을 증가시킴
- Lenti-X™ HTX Packaging Single shot 이 포함됨
HIV-1부터 유래된 lentivirus vector는 primary culture, 비분열세포, 줄기세포 및 신경세포를 포함한 거의 모든 포유류 세포에 유전자를 도입할 수 있다.
※ 본 제품은 Cloning을 위해 In-Fusion
® 시약을 함께 사용할 수 있으며, 원활한 결과를 얻기 위해
제품 사용 전 In-Fusion® 원리(동영상링크)를 반드시 참고해주시기 바랍니다.
최고 효율의 Lentivirus vector
Clontech의 모든 Lenti-X™ vector (Figure 1)는 lentivirus LTR과 packaging 서열 외에, 바이러스 역가 또는 복제 등을 크게 향상시키는 특별한 요소들을 가지고 있다.
- WPRE - Woodchuck hepatitis virus posttranscriptional regulatory element (WPRE)는 RNA의 processing과 핵외로의 이동 (nuclear export)을
촉진시키며 (1), viral genomic RNA에 작용하여 vector packaging 효율과 바이러스 역가를 높여준다. 또한, mRNA 전사를 촉진시킴으로써
목적 세포에서 실험자의 cDNA transgene의 발현을 증가시킨다.
- cPPT/CTS - Central polypyrimidine tract/central termination sequence (cPPT/CTS)는 목적 세포를 감염시키는 동안 viral genome이
핵 내로 유입 되는 것을 촉진시켜 vector의 integration과 형질도입 효율을 증가시킨다 (2).
- RRE - Rev-responsive element (RRE)는 splicing 과정을 거치지 않은 viral RNA (unspliced virus RNA)의 핵외로의 이동을 증가시켜
바이러스 역가를 향상시킨다.
Figure 1. Lenti-X™ Expression Systems and Vectors. Lenti-X™ vectors contain essential packaging sequences (lentiviral LTRs,
the Ψ packaging sequence, etc.) and additional elements that facilitate packaging and/or boost transgene expression. The Lenti-X™ Expression
System (pLVX-Puro Vector) is designed for constitutive expression of a gene of interest and provides a puromycin selectable marker. The Lenti-X™
Bicistronic Expression Systems allow your protein and a selectable marker to be coexpressed from a single mRNA transcript.
Lentiviral Packaging Systems-3가지 옵션
고역가의 Lentivirus를 제작
Lenti-X™ Packaging Single shots
Lenti-X™ Packaging Single shots은 렌티바이러스 패키징 필수 구성 유전자가 최적의 비율로 포함되어 바이러스 생산을 극대화하며,
또한 이 구성요소들이 각각의 분리된 vector로 되어 있어 바이러스 자가 복제 가능성을 효과적으로 억제하여 안정성을 확보했다.
1. VSV-G lentiviral packaging-표준의 일반적이고 가장 선호되는 Lenti-X™ Packaging Single shots (VSV-G)
(Code 631275) 은 VSV-G pseudotyped lentivirus를 제작하는데 이용되며 거의 모든 종류의 세포에 감염시킬 수 있다.
2. Ecotropic lentiviral packaging - Lenti-X™ Packaging Single shots (Ecotropic) (Code 631278) 은 MLV ecotropic envelope
glycoprotein을 가진 lentivirus를 생산하는데 이것은 mouse cell이나 rat cells에 trasnsduction시 유용하다.
3. Lenti-X™ Packaging Single shots (Integrase Deficient) (Code 631277) 은IDLV generation을 위한 4세대 lentiviral packaging system
전용으로 Lenti-X™ 293T 세포에 형질전환해서 최대 역가titer의 lentivirus를 생산할 수 있도록 최적화된 비율로 5개의 plasmid를 조합해 놓은 시스템이다.
Figure 2. Highly efficient transduction. 293T cells were infected with the indicated volumes (μl) of a Lenti-X™ supernatant
generated using the Lenti-X™ Packaging System and the pLVX-Puro vector. After puromycin selection, resistant colonies were stained with crystal violet.
Components & Storage Conditions
각 제품 구성물과 보관조건은 Certificate of Analysis 를 참조하십시오.
제품 별 vector 정보
References
1. Zufferey, R. et al. (1999) J. Virol. 73(4):2886-2892.
2. Zennou, V. et al. (2000) Cell 101(2):173-185.
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