| 型号 | 载体名称 | 出品公司 | 载体用途 |
| VRO0353 | pSUPER.retro.puro | Oligoengine | RNAi载体 |
Plasmid Type: Mammalian Expression, RNAi
Size: 6349
Bacterial Resistance: Ampicillin
Selectable Marker: Puromycin
Viral/Non-Viral: Retroviral
RNAi载体，RNAi-Ready pSIREN-Retro Q-DsRed-Express载体
| 型号 | 载体名称 | 出品公司 | 载体用途 |
| VRC0346 | RNAi-Ready pSIREN-Retro Q-DsRed-Express | Clontech | RNAi载体 |
RNAi-Ready pSIREN-RetroQ-DsRed-Express is a self-inactivating retroviral expression vector designed to express a small hairpin RNA （shRNA） using the human U6 promoter （PU6; RNA Pol III-dependent）. RNAi-Ready pSIREN-RetroQ-DsRed-Express is provided as a linearized vector digested with BamH I and EcoR I. It is used for targeted gene silencing when a ds DNA oligonucleotide encoding an appropriate shRNA is ligated into the vector. You can transfect your pSIREN-RetroQDsRed-Express construct as a plasmid expression vector, or—upon transfection into a packaging cell line—this vector can transiently express, or integrate and stably express a viral genomic transcript containing the human U6 promoter and the shRNA.
RNAi载体，RNAi-Ready pSIREN-RetroQ载体，Clontech
| 型号 | 载体名称 | 出品公司 | 载体用途 |
| VRC0345 | RNAi-Ready pSIREN-Retro Q | Clontech | RNAi载体 |
RNAi-Ready pSIREN-RetroQ is a self-inactivating retroviral expression vector designed to express a small hairpin RNA （shRNA） using the human U6 promoter （PU6; RNA Pol III-dependent）. RNAi-Ready pSIREN-RetroQ is provided as a linearized vector digested with BamH I and EcoR I. It is used for targeted gene silencing when an oligonuceotide encoding an appropriate shRNA is ligated into the vector. You can transfect your pSIREN-RetroQ construct as a plasmid expression vector, or—upon transfection into a packaging cell line—this vector can transiently express, or integrate and stably express a viral genomic transcript containing the human U6 promoter and the shRNA. The vector contains a puromycin resistance gene for the selection of stable transfectants.
| 型号 | 载体名称 | 出品公司 | 载体用途 |
| VSC0470 | pRevTet-off | Clontech | 四环素调控系统 |
pRevTet-Off™ is a retroviral vector expressing the tetracycline-controlled transactivator （tTA） from the CMV promoter. This vector is derived from pLNCX, a retroviral vector created using elements of Moloney murine leukemia virus （MoMuLV） and Moloney murine sarcoma virus （MoMuSV） as described. tTA is a fusion of amino acids 1–207 of the tet repressor （TetR） and the negatively charged C-terminal activation domain （130 amino acids） of the VP16 protein of Herpes Simplex Virus. The 5′ viral LTR controls expression of the transcript that contains Y+ （the extended viral packaging signal）, and the neomycin resistance gene （Neor） for antibiotic selection in mammalian cells. tTA is derived from vectors described previously. pRevTet-Off also includes the E. coli Ampr gene for antibiotic selection in bacteria. pRevTet-Off can be used to establish stable Tet-Off cell lines via retrovirus-mediated gene transfer. Retroviral gene transfer allows the highly efficient transduction of virtually all dividing cell types. The RevTet™ Systems are also suitable for establishing transgenic animals. In combination with the pRev-TRE retroviral expression vector, a gene of interest can be inducibly expressed at high levels in response to varying concentrations of tetracycline （Tc） or Tc derivatives such as doxycycline （Dox）. tTA binds to the Tet-response element （TRE）, thus activating transcription in the absence of Tc or Dox. As Tc or Dox is added to the culture medium, transcription from the inducible promoter is turned off in a highly dose-dependent manner. pRevTet-Off lacks the viral genes gag, pol, and env, which are supplied by the packaging cell line. It can be transfected into a high titer packaging cell line and thereby mediate production of infectious, replication-incompetent retroviral particles .The transcript produced by the pRevTet-Off construct is recognized by the viral structural proteins expressed in a packaging cell line and packaged into infectious retroviral particles. Because the RNA transcript packaged in these particles does not contain the viral genes, it cannot replicate in the target cells that it infects. The level of induction in cell populations infected with this vector depends on the efficiency of infection, the site of integration, and the titer of the virus. Viral supernatants with titers >105 cfu/ml should be produced to achieve high-level induction.
| 型号 | 载体名称 | 出品公司 | 载体用途 |
| VSC0469 | pRevTet-On | Clontech | 四环素调控系统 |
Description:
pRevTet-On is a retroviral vector expressing the reverse tetracycline-controlled transactivator
（rtTA） from the CMV promoter. This vector is derived from pLNCX, a retroviral vector created
using elements of Moloney murine leukemia virus （MoMuLV） and Moloney murine sarcoma
virus （MoMuSV） as described （1）. rtTA is a fusion of amino acids 1–207 of the reverse tet
repressor （rTetR） and the negatively charged C-terminal activation domain （130 amino acids）
of the VP16 protein of Herpes Simplex Virus. rTetR was derived from TetR and differs by four
point mutations, which are responsible for its opposite response to doxycycline. The 5′ viral
LTR controls expression of the transcript that contains Ψ+
（the extended viral packaging signal）
and the neomycin resistance gene （Neor
） for antibiotic selection in mammalian cells. rtTA is
derived from vectors described previously （2–4）. pRevTet-On also includes the E. coli Ampr
gene for antibiotic selection in bacteria.
Use:
pRevTet-On can be used to establish stable Tet-On cell lines via retrovirus-mediated gene
transfer （5）. Retroviral gene transfer allows the highly efficient transduction of virtually all
dividing cell types. The RevTet Systems are also suitable for establishing transgenic animals. In
combination with the pRev-TRE retroviral expression vector, a gene of interest can be inducibly
expressed at high levels in response to varying concentrations of the tetracycline derivative
doxycycline （Dox）. rtTA binds to the Tet-response element （TRE）, thus activating transcription in the presence of Dox. The response of rtTA to Dox is thus opposite to the response of
tTA. As Dox is removed from the culture medium, transcription from the inducible promoter
is turned off in a highly dose-dependent manner. pRevTet-On lacks the viral genes gag, pol,
and env, which are supplied by the packaging cell line. It can be transfected into a high titer
packaging cell line and thereby mediate production of infectious, replication-incompetent
retroviral particles （1, 6–7）.The transcript produced by the pRevTet-On construct is recognized
by the viral structural proteins expressed in a packaging cell line and packaged into infectious
retroviral particles. Because the RNA transcript packaged in these particles does not contain
the viral genes, it cannot replicate in the target cells that it infects.
The level of induction in cell populations infected with this vector depends on the efficiency
of infection, the site of integration, and the titer of the virus. Viral supernatants with titers >105
cfu/ml should be produced to achieve high-level induction.