pSingle-tTS-shRNA



 

编号

载体名称

北京华越洋VECT76562

pSingle-tTS-shRNA

 

pSingle-tTS-shRNANA载体基本信息:

质粒类型:

RNAi载体

克隆方法:

多克隆位点,限制性内切酶

载体大小:

6990 bp

3' 测序引物:

Sing-Tet-U6-R: (4999–4967): 5'-GAAGC GGAAG AGCGC CCAAT ACGCA

 AACCG CCT-3'

载体抗性:

Ampicillin (氨苄青霉素)

筛选标记:

Neomycin (新霉素)

 

pSingle-tTS-shRNA载体图谱:

pSingle-tTS-shRNA载体简介:

 

The pSingle-tTS-shRNA vector expresses the tetracyline-controlled transcriptional suppressor (tTS), which controls expression of an shRNA sequence inserted into the shRNA cloning site. The tTS protein is a fusion of the Tet repressor protein (TetR) and the KRAB-AB silencing domain of the Kid-1 protein (SDKid-1 ), a powerful transcriptional suppressor (1, 2). In the absence of the inducer doxycycline (Dox, a tetracycline derivative), tTS binds to the tetO sequences in the modified Tet-responsive Pol III hybrid promoter (P Tight/U6 ) and blocks expression of the shRNA. As Dox is added to the culture medium, tTS dissociates from P Tight/U6 to allow Pol IIImediated transcription of the shRNA, resulting in suppression of the target gene in a highly dose-dependent manner. pSingle-tTS-shRNA also contains a ColE1 origin of replication and an ampicillin resistance gene (Amp r ) for propagation and selection in bacteria, as well as a neomycin resistance gene (Neo r ) for selection of stable transformants in mammalian cells.

pSingle-tTS-shRNA载体应用

After digesting pSingle-tTS-shRNA with HindIIIand XhoI, clone an annealed ds oligonucleotide encoding your desired shRNA sequence into the vector. Transfect the recombinant vector into cells and select stable, clonal tranformants with G418 to obtain a cell line capable of induced suppression of the shRNA's target gene.

 

pSingle-tTS-shRNA载体相关的RNAi载体:

pTRIPZ controlpSilencer 3.1-H1 neo
pSUPER.puropSilencer 3.1-H1 hygro
pSUPER.neo+gfppSilencer 3.0-H1
pSUPERpSilencer 2.1-U6 hygro
psiRNA-hH1neo G2pRNAT-U6.1/neo
psiRNAH1-neopRNATin-H1.2/Retro
pSingle-tTS-shRNApRNATin-H1.2/Neo
pSilencer adeno 1.0-CMVpRNATin-H1.2/Hygro
pSilencer 4.1-CMV puropRNA-H1.1/Adenoe
pSilencer 4.1-CMV neopmR-mCherry
pMIR-REPORT LuciferasepGIPZ empty
pLVTHpDC316-ZsGreen-shRNA


 


 

 

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