Lentiviral CRISPR/Cas9 System
Lentiviral CRISPR/Cas9 System
Brand: Takara Bio.
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Lentiviral CRISPR/Cas9 System
The Lenti-X CRISPR/Cas9 System and Lenti-X Tet-On 3G CRISPR/Cas9 System are complete systems for lentiviral-mediated CRISPR/Cas9 genome editing. By using lentivirus to deliver sgRNA and Cas9, these systems allow you to achieve targeted genome editing in cell lines that are difficult to transfect. Each system includes Lenti-X Packaging Single Shots for generating high titers of virus and plasmid sets designed to express Cas9 and your custom sgRNA in target cells. The plasmid for sgRNA expression, pLVX-hyg-sgRNA1, comes prelinearized for easy insertion of your sgRNA sequence and is supplied in sufficient quantity for the construction of 10 different plasmids, each with its own sgRNA sequence.
Whereas the Lenti-X CRISPR/Cas9 System involves the constitutive expression of Cas9 in target cells, the Lenti-X Tet-On 3G CRISPR/Cas9 System incorporates the Tet-On 3G transactivator protein, which enables users to induce Cas9 expression via the addition of doxycycline. By allowing for tight control of Cas9 expression, the Lenti-X Tet-On 3G CRISPR/Cas9 System enables users to minimize the likelihood of toxicity and off-target effects associated with persistent Cas9 expression in cell culture.
Overview
- Viral delivery of the Cas9 gene and a sgRNA allows for genome editing in hard-to-transfect mammalian cells, including proliferating and non-proliferating cells
- Lenti-X Packaging Single Shots make generating high-titer lentivirus extremely simple
- Separate systems available for either constitutive or inducible Cas9 expression in target cells
- Tightest possible control of Cas9 expression (Lenti-X Tet-On 3G CRISPR/Cas9 System) allows you to minimize the likelihood of toxicity and off-target effects associated with persistent Cas9 expression in cell culture
- Learn about phenotypic screening using a lentiviral whole genome CRISPR library.
Applications
- Lentivirus-based delivery of a user-defined sgRNA and Cas9 for mammalian genome editing using CRISPR/Cas9 technology
Either Jurkat or HT1080 cells were first transduced with LVX-hyg-CD81-sgRNA and then selected for stable integration using hygromycin
Either Jurkat or HT1080 cells were first transduced with LVX-hyg-CD81-sgRNA and then selected for stable integration using hygromycin. Stable clones were then transduced with LVX-puro-Cas9. Stable clones were selected for using puromycin and then screened for CD81 knockout efficiency using FACS. Positive and negative controls of parental cells without Cas9 transduction were done either with or without anti-CD81 antibody.
Components
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Lenti-X CRISPR/Cas9 System (Cat. # 632629)
- pLVX-hyg-sgRNA1 Vector (Linear)
- pLVX-puro-Cas9 Vector
- Stellar Competent Cells
- Guide-it Ligation Components v2
- Lenti-X Packaging Single Shots
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Lenti-X Tet-On 3G CRISPR/Cas9 System (Cat. # 632633)
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pLVX-hyg-sgRNA1 Vector (Linear)
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pLVX-TRE3G-Cas9-puro Vector Set
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pLVX-EF1a-Tet3G Vector
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Stellar Competent Cells
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Guide-it Ligation Components v2
-
Lenti-X Packaging Single Shots
-
-
pLVX-hyg-sgRNA1 Vector System (Cat. # 632630)
-
pLVX-hyg-sgRNA1 Vector (Linear)
-
Guide-it Ligation Components v2
-
Stellar Competent Cells
-
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Either Jurkat or HT1080 cells were first transduced with LVX-hyg-CD81-sgRNA and then selected for stable integration using hygromycin
Either Jurkat or HT1080 cells were first transduced with LVX-hyg-CD81-sgRNA and then selected for stable integration using hygromycin. Stable clones were then transduced with LVX-puro-Cas9. Stable clones were selected for using puromycin and then screened for CD81 knockout efficiency using FACS. Positive and negative controls of parental cells without Cas9 transduction were done either with or without anti-CD81 antibody.
Components
-
Lenti-X CRISPR/Cas9 System (Cat. # 632629)
- pLVX-hyg-sgRNA1 Vector (Linear)
- pLVX-puro-Cas9 Vector
- Stellar Competent Cells
- Guide-it Ligation Components v2
- Lenti-X Packaging Single Shots
-
Lenti-X Tet-On 3G CRISPR/Cas9 System (Cat. # 632633)
-
pLVX-hyg-sgRNA1 Vector (Linear)
-
pLVX-TRE3G-Cas9-puro Vector Set
-
pLVX-EF1a-Tet3G Vector
-
Stellar Competent Cells
-
Guide-it Ligation Components v2
-
Lenti-X Packaging Single Shots
-
-
pLVX-hyg-sgRNA1 Vector System (Cat. # 632630)
-
pLVX-hyg-sgRNA1 Vector (Linear)
-
Guide-it Ligation Components v2
-
Stellar Competent Cells
-
