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产品货号 英文简称 产品名称 规格 价格
PiggyBac Gene Editing HR Targeting Vector (MCS1-5'PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3' PB TR-MCS2) PiggyBac Gene Editing HR Targeting Vector (MCS1-5'PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3' PB TR-MCS2) PiggyBac Gene Editing HR Targeting Vector (MCS1-5'PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3' PB TR-MCS2) 10ug
产品说明

Without a trace—make seamless gene edits with no residual footprint—includes dual GFP/puromycin selection and on-target enrichment with TK selection

Overview

Seamless gene editing

Use the PiggyBac Gene Editing HR Targeting Vector (MCS1-5’PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3′ PB TR-MCS2) to get seamless gene editing—leave no trace of vector sequences behind.

 

This special HR Donor works with the  instead of the Cre-LoxP system. Simply proceed with your CRISPR/Cas9 gene editing as usual—clone your homology arms into MCS1 and MCS2, use dual GFP and puromycin selection to find integrants, and enrich for on-target events using negative thymidine kinase (TK) selection (Figure 1).

 

Why use an HR targeting vector?

Even though gene knock-outs can result from DSBs caused by Cas9 alone, SBI recommends the use of HR targeting vectors (also called HR donor vectors) for more efficient and precise mutation。 HR donors can supply elements for positive or negative selection ensuring easier identification of successful mutation events。 In addition, HR donors can include up to 6-8 kb of open reading frame for gene knock-ins or tagging, and, when small mutations are included in either 5’ or 3’ homology arms, can make specific, targeted gene edits。

Choose the right HR Targeting Vector for your project

Catalog # HR Donor Vector Features* Application
GENE KNOCK-OUT GENE KNOCK-IN GENE EDITS GENE TAGGING
HR100PA-1 Basic HR Donor    
HR110PA-1 Removable RFP marker and puromycin selection    
HR120PA-1 Tag with GFP fusion 
Removable RFP marker and puromycin selection
     
HR130PA-1 Co-express GFP with “tagged” gene via T2A 
Removable RFP marker and puromycin selection
     
HR150PA-1 Tag with GFP fusion and co-express luciferase via T2A 
Removable RFP marker and puromycin selection
     
HR180PA-1 Co-express GFP with “tagged” gene via IRES 
Removable RFP marker and puromycin selection
     
HR210PA-1 Removable GFP marker, puromycin selection, and TK selection    
HR220PA-1 Tag with GFP fusion 
Removable RFP ,arker and hygromycin Selection
     
HR410PA-1 Removable GFP marker and puromycin selection    
HR510PA-1 Removable RFP marker and hygromycin selection        
HR700PA-1 Enrich for on-target integration with negative TK selection** 
Removable GFP marker and puromycin selection
   
HR710PA-1 Enrich for on-target integration with negative TK selection** 
Removable RFP marker and hygromycin selection
   
HR720PA-1 Enrich for on-target integration with negative TK selection** 
Removable blasticidin selection
   
GE602A-1 First generation AAVS1-targeting HR Donor      
GE603A-1 First generation AAVS1-targeting HR Donor (positive control for GE602A-1)      
GE620A-1 Second generation AAVS1-targeting HR Donor 
Promoterless to knock-in any gene or promoter-gene combination
     
GE622A-1 Second generation AAVS1-targeting HR Donor 
Constitutive expression of your gene-of-interest
     
GE624A-1 Second generation AAVS1-targeting HR Donor 
Create reporter cell lines
     
CAS620A-1 Knock-in Cas9 to the AAVS1 site        
PBHR100A-1 Use with the PiggyBac Transposon System 
Enables seamless gene editing with no residual footprint (i.e. completely remove vector sequences)
     
*All HR Target Vectors except PBHR100A-1 contain LoxP sites. Any sequences that are integrated between the two LoxP sites can be removed through transient expression of Cre Recombinase. 
**The clever design of these HR Donors enables enrichment for on-target integration events. A PGK-hsvTK cassette is included outside of the homology arms. Because of this configuration, on-target integration that results from homologous recombination will not include the PGK-hsvTK cassette—only randomly-integrated off-target events will lead to integration of PGK-hsvTK and resulting TK activity. Therefore, TK selection will negatively select against off-target integrants. Click on any one of these vectors to see a diagram of how the negative selection works.
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