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Lentiviral expression vector를 포함한 시스템 - 초기 세팅에 최적

CMV promoter 벡터 시스템

제조사	제품코드	제품명	용량	가격 (부가세별도)	사용자매뉴얼
Clontech	631253	Lenti-X™ Expression System (EF1a Version)	set (Package)	2,225,000원	, ,
Clontech	632164	Lenti-X™ Expression System	1 Set (Package)	2,225,000원	, ,
Clontech	632181	Lenti-X™ Bicistronic Expression System (Neo)	1 Kit (Package)	2,225,000원	, ,
Clontech	632182	Lenti-X™ Bicistronic Expression System (Hyg)	1 Kit (Package)	2,225,000원	, ,
Clontech	632183	Lenti-X™ Bicistronic Expression System (Puro)	1 Kit (Package)	2,225,000원	, ,
Clontech	631275	Lenti-X™ Packaging Single Shots (VSV-G)	16 회	1,460,000원
Clontech	631276	Lenti-X™ Packaging Single Shots (VSV-G)	96 회	4,278,000원
Clontech	631277	Lenti-X™ Packaging Single Shots (Integrase Deficient)	16 회	1,878,000원
Clontech	631278	Lenti-X™ Packaging Single Shots (Ecotropic)	16 회	1,878,000원

Product index

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.

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 pacakaging 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

Vector명	정보
pLVX-Puro Vector (#632164 구성품)	Vector map	Sequence (txt.)
pLVX-EF1alpha-IRES-Puro Vector (#631253 구성품)	Vector map	Sequence (txt.)
pLVX-IRES-Neo vector (#632181 구성품)	Vector map	Sequence (txt.)
pLVX-IRES-Hyg Vector (#632182 구성품)	Vector map	Sequence (txt.)
pLVX-IRES-Puro Vector (#632183 구성품)	Vector map	Sequence (txt.)

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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Keyword :

Lentivirus

[개요] Packaging Single Shots

[FAQ] Lentiviral Gene Delivery

Lentiviral expression vector를 포함한 시스템..

CMV promoter 벡터 시스템

조혈모세포, 줄기세포 등의 Lentiviral transdu..

EF1 α promoter 벡터 시스템

SARS-CoV-2-pseudotyped lentivirus 생산

Lenti-X™ SARS-CoV-2 Packaging Mix

임상 치료제 개발을 위한

3세대 Lentivirus packaging system

다양한 형광단백질 리포터를 포함한 Lentiviral..

형광단백질 리포터 벡터

Tetracycline 존재 하에서만 목적 단백질을 발..

Tet-on inducible Expression (Two vectors)

Tetracycline 존재 하에서만 목적 단백질을 발..

Tet One Inducible Expression (One vector)

Lentivirus를 이용한 sgRNA, Cas9 gene deliver..

Lenti-X™ CRISPR/Cas9 System

Tetracycline 존재 하에서만 목적 단백질을 발..

(Legacy) Tet-Inducible expression 시스템

Pormoterless vector로 pomorter 활성 연구

Reporter assay 시스템 (Promoterless vector)

Transfection 시약을 포함하여 더 간단한 packa..

Packaging single shots

239T, HEK-293 Cell보다 더 높은 packaging 효..

Packaging Cell Lines

1 step qRT-PCR로 역가를 빠르고 정확하게 측정

Lenti-X™ qRT-PCR Titration Kit

p24 ELISA로 빠르고 정확하게 역가 측정

Lenti-X™ p24 Rapid Titer Kit (Single Wash)

스마트폰으로 10분만에 Lentivirus 역가(Titer)..

GoStix™ Plus

Lentivirus 농도가 낮을 때, Mix & Spin으로 간..

Lentivirus 농축 시약

In vivo Lentiviral delivery 적용 시, 필수적..

Lentivirus 정제

Ecotropic lentivirus를 human cell에 적용

Ecotropic Receptor Booster

Lentivirus나 MMLV retrovirus의 더 빠른 trans..

Transduction 시약

목적 세포의 유전자에 삽입된 Provirus copy nu..

Provirus Quantitation Kit

Lentivirus가 목적세포 유전자에 삽입된 위치를..

Integration Site Analysis Kit

최적화된 Transfection Reagent & Packaging Mi..

TransIT® Lentivirus System