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System Biosciences

Basic HR Targeting Vector [MCS1-LoxP-MCS2-MCS3-pA-LoxP-MCS4] for Gene Knock-In/Out

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SKU:
HR100PA-1
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10 ug
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  • Basic HR Targeting Vector [MCS1-LoxP-MCS2-MCS3-pA-LoxP-MCS4] for Gene Knock-In/Out
  • Basic HR Targeting Vector [MCS1-LoxP-MCS2-MCS3-pA-LoxP-MCS4] for Gene Knock-In/Out
£880.64

Description

Basic HR Targeting Vector [MCS1-LoxP-MCS2-MCS3-pA-LoxP-MCS4] for Gene Knock-In/Out . Cat# HR100PA. Supplier: SBI System Biosciences

Knock-in any expression cassette to a specific genomic location with this basic HR targeting vector

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Overview

Get precise genomic integration of your expression cassette 
 
Use the PrecisionX™ Basic HR Targeting Vector for Gene Knock-In/Out (MCS1-LoxP-MCS2-MCS3-pA-LoxP-MCS4) to insert any expression cassette into a specific location of the genome for both gene knock-ins—i.e. to express a gene-of-interest from a specific location such as the AAVS1 Safe Harbor Site—and gene knock-outs—i.e. to knock-out a gene by inserting specific sequences such as a GFP reporter cassette.
 
The Basic HR Targeting Vector for Gene Knock-In/Out (MCS1-LoxP-MCS2-MCS3-pA-LoxP-MCS4) features four different MCSs—clone your homology arms into MCS1 and MCS4, your expression cassette into MCS3, and any additional sequences such as small RNAs that don’t need a polyA tail into MCS2—as well as two LoxP sites that can be used to remove the expression cassette after it is no longer needed (learn more about Cre-LoxP excision here).
 
 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

How It Works

At-a-glance—how to use an HR Targeting Vector to knock-in a gene

Figure 1. Knocking-in a gene using an HR Targeting Vector. 

Step 1: Cas9 creates a double-stranded break(DSB) in the genomic DNA at a site that is complimentary to the gRNA. 

Step 2: The DNA repair machinery is recruited to the DSB. In the presence of an HR Donor with homology to the region adjacent to the DSB (blue areas of the genomic and vector DNA) homologous recombination (HR) is favored over non-homologous end joining (NHEJ).

 Result: The HR event leads to insertion of the region of the HR Donor Vector between the two homology arms—your gene-of-interest is integrated into the genome.

At-a-glance—how to use an HR Targeting Vector to knock-out a gene

Figure 2. Knocking-out a gene using an HR Targeting Vector.

 Step 1: Cas9 creates a double-stranded break(DSB) in the genomic DNA at a site that is complimentary to the gRNA.

 Step 2: The DNA repair machinery is recruited to the DSB. In the presence of an HR Donor with homology to the region adjacent to the DSB (blue areas of the genomic and vector DNA) homologous recombination (HR) is favored over non-homologous end joining (NHEJ).

 Result: The HR event leads to insertion of the region of the HR Donor Vector between the two homology arms—your selection cassette is integrated into the gene, disrupting the open reading frame.

 

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