Cas9 Stable Cell Line - SH-SY5Y
Cat.No. : CSC-RO0181 Host Cell: SH-SY5Y
Size: >1x10^6 cells/vial Validation: T7 Endonuclease I assay
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Cell Line Information
Safety and Packaging
| Cat. No. | CSC-RO0181 |
| Product Type | Cas9 overexpression stable cell line |
| Introduction | Clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 is a gene-editing technology that contains two essential components: a guide RNA (gRNA) to match a target gene, and the Cas9 (CRISPR-associated protein 9) endonuclease which causes a double-stranded DNA break, allowing modifications to the genome via nonhomologous end joining (NHEJ) or homology-directed repair (HDR). |
| Cell Line Information | SH-SY5Y-Cas9 cell line is engineered to stably overexpress Cas9 nuclease. The Cas9 nuclease in SH-SY5Y-Cas9 cell line has been functionally validated using T7 Endonuclease I assay. In combination with separately transfected sgRNAs, SH-SY5Y-Cas9 cell line can be used to efficiently generate targeted genomic modifications including gene knockout, gene knockin, gene mutagenesis, gene tagging etc. It is also an ideal cell line model for sgRNA screening and validation, either individually or in pools. |
| Target Gene | Cas9 |
| Host Cell | SH-SY5Y |
| Applications | 1) CRISPR genome editing, such as gene knockout (KO), gene knockin (KI), gene mutagenesis, gene tagging etc. 2) High-throughput sgRNA screening and validation |
| Quality Control | 1) T7E1 assay 2) Mycoplasma detection |
| Size Form | One vial of frozen cells, typically >1x10^6 cells/vial |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
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Background
Case Study
Applications
Cas9 is a groundbreaking molecular tool in the field of genetic engineering, discovered in 2012 by Jennifer Doudna and Emmanuelle Charpentier. It is an RNA-guided DNA endonuclease derived from the bacterial CRISPR-Cas immune system. Cas9 enables precise genome editing by inducing double-strand breaks at specific target sites within the DNA sequence. This technology revolutionized gene editing due to its simplicity, efficiency, and versatility.
SY5Y, a human neuroblastoma cell line, has been extensively employed in neuroscience research. Originally established in 1970, SY5Y cells exhibit characteristics reminiscent of immature sympathetic neurons, making them a valuable model for studying neuronal development, differentiation, and neurodegenerative diseases.
The development of Cas9 Stable Cell Line-SH-SY5Y integrates the power of Cas9 technology with the utility of the SY5Y cell line. This engineered cell line, established in recent years, facilitates efficient and precise genome editing within the SY5Y cellular context. It serves as a potent tool for elucidating the molecular mechanisms underlying neurodevelopmental processes and neurological disorders, offering unprecedented opportunities for advancing our understanding of neuronal biology.
Researchers employed CRISPR‐Cas9 technology to investigate the impact of a rare mutation (rs144662445) in the AKAP9 gene, linked to Alzheimer's disease (AD) in African Americans (AA), on tau pathology and the tau interactome in SH‐SY5Y P301L neuron‐like cells. The mutation notably elevated phosphorylated tau levels, particularly at Ser396/Ser404. Additionally, analysis of the tau interactome revealed dysregulation of RNA translation, localization, and oxidative activity, resembling findings from human AD brain samples. Functional assays confirmed reduced protein synthesis and increased oxidative stress, mimicking AD pathology. These findings highlight the role of AKAP9 in AD pathogenesis.
Figure 1. CRISPR-mediated AKAP9 I2558M mutation knock-in was performed in SH-SY5Y P301L cells, followed by neuronal differentiation. Mutation generation was confirmed by Sanger sequencing. (You Y, et al., 2022)
Creative Biogene's Cas9 Stable Cell Line-SH-SY5Y expedites similar studies, eliminating the need for gene editing steps. This pre-engineered cell line directly investigates gene mutation effects on tau pathology and the tau interactome, saving time and costs while accelerating disease mechanism understanding.
1. Gene Editing: Utilize CRISPR/Cas9 system to knockout or knock-in specific genes in SH-SY5Y cells, enabling the study of gene function in neurobiology.
2. Disease Modeling: Introduce disease-associated mutations into SH-SY5Y cells to mimic pathological conditions such as Parkinson's or Alzheimer's disease, facilitating mechanistic studies.
3. Drug Screening: Employ CRISPR/Cas9-modified SH-SY5Y cells to screen potential therapeutics for neurodegenerative disorders, enhancing drug discovery efforts.
4. Signal Transduction Studies: Investigate signaling pathways involved in neuronal development and function by manipulating target genes in SH-SY5Y cells.
5. Neurotoxicity Assessment: Assess the impact of environmental toxins or pharmaceutical compounds on neuronal viability and function using genetically engineered SH-SY5Y cell lines.
For research use only. Not intended for any clinical use.
