pHAGE TRE dCas9-KRAB BioVector?哺乳動(dòng)物慢病毒基因編輯質(zhì)粒-BioVector NTCC質(zhì)粒載體菌株細(xì)胞蛋白抗體基因保藏中心

BioVector? pHAGE TRE dCas9-KRAB

Understanding the Components:

• pHAGE: This likely refers to a lentiviral vector system, a common tool for delivering genetic material into cells. Lentiviruses can integrate their genetic material into the host cell's genome, allowing for stable and long-term expression.

• TRE: This stands for Tetracycline Response Element. It's a DNA sequence that can be activated or repressed by the presence or absence of tetracycline or its derivatives, such as doxycycline. This allows for inducible gene expression.

• dCas9: A deactivated form of the Cas9 protein. While it can still bind to specific DNA sequences guided by a single-guide RNA (sgRNA), it lacks the ability to cut DNA.

• KRAB: A transcriptional repressor domain. When fused to dCas9, it can effectively silence gene expression by binding to the target DNA sequence and recruiting other proteins to repress transcription.

How it Works:

1. Lentiviral Delivery: The pHAGE vector containing the TRE-dCas9-KRAB construct is packaged into lentiviral particles.

2. Cell Infection: These particles infect target cells, and the viral DNA integrates into the host genome.

3. Inducible Expression: In the absence of doxycycline, the TRE promoter is inactive, and dCas9-KRAB is not expressed.

4. Gene Repression: When doxycycline is added, it binds to a regulatory protein, activating the TRE promoter. This leads to the expression of dCas9-KRAB.

5. Target Gene Silencing: The dCas9-KRAB complex binds to specific DNA sequences targeted by sgRNAs, leading to transcriptional repression of the target gene.

Applications:

• Gene Function Studies: Silencing genes to study their role in cellular processes.

• Disease Modeling: Creating cellular models of diseases by repressing specific genes involved in disease pathogenesis.

• Therapeutic Development: Identifying potential drug targets by silencing genes and observing phenotypic effects.

• Genome Engineering: Precisely controlling gene expression in a temporal and spatial manner.

Key Advantages:

• Inducible Expression: Allows for precise control of gene repression.

• High Specificity: The CRISPR-Cas9 system enables highly specific targeting of genes.

• Potent Repression: The KRAB domain is a powerful transcriptional repressor.

• Versatile Platform: Can be used in a variety of cell types and organisms.

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BioVector NTCC質(zhì)粒載體菌株細(xì)胞蛋白抗體基因保藏中心

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