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Creative Biogene's SuperKO sgRNA Clones leverage sgRNA's vital role in CRISPR/Cas9, enabling precise gene knockout in prokaryotes. SuperKO sgRNA Clones, designed specifically for gene knockout, are CRISPR/Cas9-engineered clones intended to facilitate efficient gene knockout experiments for researchers. These SuperKO sgRNA Clones not only facilitate gene knockout experiments for researchers but also accelerate progress in gene function studies and drug target screening in various application domains.
Key Features of Our SuperKO sgRNA Clones
sgRNA, as a crucial component of the CRISPR/Cas9 system, plays a pivotal role in gene knockout. Comprising a 20-nucleotide target-specific sequence and a structural RNA element derived from Streptococcus pyogenes tracrRNA, sgRNA enables precise targeting of any sequence in the genome. The process of Cas9/sgRNA complex binding to the target site and subsequent DNA cleavage is multi-step, often resulting in small insertion and deletion (indel) mutations at the cleavage site upon repair via the non-homologous end joining (NHEJ) pathway, thereby silencing gene expression. Specifically, the following tasks can be performed:
Case Study 1
Researchers aimed to elucidate the role of MDM2 E3 ligase activity in regulating p53 and cell cycle progression. They utilized an MDM2 sgRNA Vector to specifically disrupt MDM2 E3 ligase function while maintaining its interaction with MDM4. This allowed them to investigate the impact of MDM2 E3 ligase activity on p53 regulation in vivo. The study revealed that Mdm2L466A/L466A mice, lacking MDM2 E3 ligase activity, exhibited embryonic lethality due to p53-dependent mechanisms, highlighting the essential role of MDM2 E3 ligase in p53 regulation. Additionally, they observed unexpected cell cycle defects and increased aneuploidy in cells expressing the E3-dead MDM2 mutant, suggesting a p53-independent role of MDM2 E3 ligase in cell cycle regulation and genome integrity maintenance. These findings shed light on the multifaceted functions of MDM2 in cellular processes beyond its canonical role in p53 regulation.
Figure 1. Researchers analyzed p53 expression using WB analysis in E9.5 embryos of different genotypes. They utilized sgRNA vector to knock out the target gene and observed normalized p53 levels. Additionally, they performed WB analysis in MEFs with different treatments to study p53, MDM2, and MDM4 protein expression levels. (Chinnam M, et al., 2022)
Case Study 2
The effectiveness of BET protein inhibitors (BETi's) in treating acute myeloid leukemia (AML) is hindered by the development of adaptive or innate resistance. Researchers used sgRNA vectors to knock out specific genes and understand BET protein inhibitor (BETi) resistance in acute myeloid leukemia (AML). They found increased activity in certain chromatin regions associated with β-catenin, TCF7L2, JMJD6, and c-Myc in resistant AML cells. Knocking out TCF7L2 or JMJD6 reversed resistance, while overexpression conferred resistance. Patient-derived AML cells with resistance also showed elevated expression of these genes. Targeting them with CRISPR/Cas9 reduced cell viability. Combining small-molecule inhibitors with BETi's reduced c-Myc levels and improved survival in mouse models. This suggests targeting multiple components of the resistance mechanism can overcome BETi resistance in AML cells.
Figure 2. The target genes' expression was depleted, cell viability was reduced, and sensitivity to BET inhibitor treatment was restored in SET-2 and SET-2-OTX P/R cells through the knockout of TCF7L2 and JMJD6 using sgRNA vectors. Successful knockout and phenotypic changes were confirmed by immunoblot and confocal microscopy analyses. (Saenz DT, et al., 2020)
Q: How does the quality of the SuperKO sgRNA Clones ensure reliable results in genome editing?
A: The SuperKO sgRNA Clones are meticulously designed and validated to ensure high efficiency and specificity in gene knockout. Each sgRNA undergoes rigorous quality control measures to guarantee accuracy and reproducibility in genome editing experiments.
Q: What measures are in place to address potential off-target effects associated with CRISPR/Cas9 editing?
A: To mitigate off-target effects, the sgRNA sequences are carefully selected and screened to minimize potential binding to unintended genomic loci. Additionally, experimental protocols are optimized to enhance the specificity of CRISPR/Cas9 editing.
Q: How do the SuperKO sgRNA Clones streamline the process of multiplex editing?
A: The SuperKO sgRNA Clones allow for simultaneous editing of multiple genes without the need for marker insertion, thereby simplifying the experimental workflow and saving time and resources in genome engineering endeavors.
Q: Can the SuperKO sgRNA Clones be customized for specific gene targets?
A: Yes, the SuperKO sgRNA Clones can be tailored to target specific genes of interest, providing flexibility and versatility for various research applications.
Q: How does the understanding of sgRNA activity contribute to the development of novel antimicrobials?
A: Insight into sgRNA activity aids in designing antimicrobial agents that effectively target bacterial cells, thereby offering potential solutions to combat antibiotic resistance and infectious diseases.
| Cat.No. | Product Name | Price |
|---|---|---|
| SKO0001 | CALD1 Validated sgRNA vector | Inquiry |
| SKO0002 | ALPP Validated sgRNA vector | Inquiry |
| SKO0003 | MVP Validated sgRNA vector | Inquiry |
| SKO0004 | ABCC3 Validated sgRNA vector | Inquiry |
| SKO0005 | TFE3 Validated sgRNA vector | Inquiry |
| SKO0006 | CD44 Validated sgRNA vector | Inquiry |
| SKO0007 | MDM2 Validated sgRNA vector | Inquiry |
| SKO0008 | HIF1A Validated sgRNA vector | Inquiry |
| SKO0009 | KRAS Validated sgRNA vector | Inquiry |
| SKO0010 | COL4A1 Validated sgRNA vector | Inquiry |
| SKO0011 | LAMA1 Validated sgRNA vector | Inquiry |
| SKO0012 | CASP3 Validated sgRNA vector | Inquiry |
| SKO0013 | S100A4 Validated sgRNA vector | Inquiry |
| SKO0014 | CD9 Validated sgRNA vector | Inquiry |
| SKO0015 | TNFRSF10A Validated sgRNA vector | Inquiry |
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