sgBbsI(p2ToI-U6-2xBbsI-SgRNA-HygR) BioVector® Tol2转座子基因编辑质粒 BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

sgBbsI(p2ToI-U6-2xBbsI-SgRNA-HygR) BioVector® Tol2转座子基因编辑质粒

BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

sgBbsI(p2ToI-U6-2xBbsI-SgRNA-HygR) is a plasmid designed for CRISPR-Cas9-mediated genome editing.

Here's a breakdown of its components:

• sgBbsI: This likely refers to the BbsI restriction enzyme recognition site within the plasmid. BbsI is commonly used for cloning single-guide RNA (sgRNA) sequences into CRISPR plasmids.

• p2ToI-U6-2xBbsI-SgRNA-HygR: This part of the plasmid name describes its key features:

  • p2ToI: This may indicate the plasmid backbone or origin.

  • U6:  The U6 promoter is a strong RNA polymerase III promoter used to drive the expression of the sgRNA.

  • 2xBbsI: This suggests the presence of two BbsI restriction enzyme sites flanking the sgRNA cloning region.

  • SgRNA: This represents the region where the user will insert the specific 20-nucleotide sequence targeting the desired genomic locus.

  • HygR: This gene confers resistance to the antibiotic hygromycin, allowing for the selection of cells that have successfully integrated the plasmid.

  
  

How it Works:

1. sgRNA Design: You design a 20-nucleotide sgRNA sequence that specifically targets the gene of interest.

2. Cloning: The sgRNA sequence is synthesized as two complementary oligonucleotides containing BbsI overhangs. These oligonucleotides are then annealed and ligated into the BbsI-digested plasmid.

3. Transformation: The plasmid containing the sgRNA is transformed into the target cells.

4. Selection: Cells that have successfully integrated the plasmid are selected by growing them in the presence of hygromycin.

5. Genome Editing: The expressed sgRNA guides the Cas9 protein to the target genomic locus, leading to DNA cleavage and subsequent gene editing events (e.g., gene knockout, gene insertion).

Key Applications:

• Gene Knockout: Disrupting gene function by inducing indels (insertions or deletions) at the target site.

• Gene Editing: Introducing specific mutations or modifications into the target gene.

• Functional Genomics: Studying gene function by analyzing the effects of gene disruption.

In summary: sgBbsI(p2ToI-U6-2xBbsI-SgRNA-HygR) is a valuable tool for researchers conducting CRISPR-Cas9-mediated genome editing experiments in various organisms.

Map

生产厂家Supplier:

BioVector NTCC质粒载体菌株细胞蛋白抗体基因保藏中心

E-mail:BioVector@163.com

http://www.biovector.net