Accelerate research, empower industry
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Overview Cas9 cell line gene mutation (CGM) stands as a highly effective tool for investigating cell gene functions, particularly enabling the in-situ study of point mutations within cells. Key Advancements
Our achievements With a track record of over 100 successful cases, Hysigen has demonstrated proficiency in transfecting commonly used cell lines, including Jurkat, NK-92, BV2, C2C12, EMT6, B16-F10, ARPE-19, HepG2, THP-1, HCT116, A549, RAW264.7, MDA-MB-231, MDA-MB-468, 4T1, and more. Workflow
* We kindly remind you that we provide gene editing services for primary cells, stem cells, or iPS cells.
 CRISPR-Mediated Point Mutation CRISPR-mediated point mutation involves using the CRISPR-Cas9 system to introduce precise changes in a single nucleotide of a targeted gene. The process begins with the design of a guide RNA (gRNA) that directs the Cas9 enzyme to the specific genomic location where the point mutation is desired. The Cas9 protein acts as molecular scissors, creating a double-strand break (DSB) at the target site. The cell's natural repair mechanism, Homology Directed Repair (HDR), then comes into play. HDR can be employed with a donor DNA template containing the desired point mutation. This template guides the precise repair of the DNA, incorporating the desired mutation at the targeted nucleotide position. The CRISPR-mediated point mutation mechanism enables researchers to engineer specific changes in the genome with remarkable precision. Point mutation of JAK2 in MEG-01 Cells Using CRISPR/Cas9
• Design & Transfection: The gRNA and ssDNA donor template were designed to target the specific region of the JAK2 gene and introduce the V617F mutation (Fig. 1). They were co-electroporated into MEG-01 cells. • Single-Cell Cloning: Single cells were isolated using ClonePlus™ technology to ensure monoclonality. • Screening & Expansion: Monoclonal populations in 96-well plates were screened via Sanger sequencing (Fig. 2). The confirmed positive clone with the V617F mutation was expanded for further analysis. Figure 1. JAK2 V617F targeting strategy.
Figure 2. Genotypic validation of the JAK2 V617F mutation MEG-01 cell line by Sanger sequencing.
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IF: 45.5 Chen Y, Chen S, Liu Z, Wang Y, An N, Chen Y, Peng Y, Liu Z, Liu Q, Hu X. Red blood cells undergo lytic programmed cell death involving NLRP3. Cell. 2025 Apr 16:S0092-8674(25)00389-7. | IF: 39.3 Ma B, Ju A, Zhang S, et al. Albumosomes formed by cytoplasmic pre-folding albumin maintain mitochondrial homeostasis and inhibit nonalcoholic fatty liver disease[J]. Signal Transduction and Targeted Therapy, 2023, 8(1): 229. | IF: 26.6 Wu W, Pu Y, Gao S, et al. Bacterial Metabolism-Initiated Nanocatalytic Tumor Immunotherapy[J]. Nano-Micro Letters, 2022, 14(1): 1-21. | IF: 37.3 Zheng Z, Zeng X, Zhu Y, et al. CircPPAP2B controls metastasis of clear cell renal cell carcinoma via HNRNPC-dependent alternative splicing and targeting the miR-182- 5p/CYP1B1 axis[J]. Molecular Cancer, 2024, 23(1): 4. | IF: 18.9 Sun J, Yang F, Wang L, et al. Delivery of coenzyme Q10 loaded micelle targets mitochondrial ROS and enhances efficiency of mesenchymal stem cell therapy in intervertebral disc degeneration[J]. Bioactive Materials, 2023, 23: 247-260. | IF: 18.9 Wei X, Wang L, Duan C, et al. Cardiac patches made of brown adipose-derived stem cell sheets and conductive electrospun nanofibers restore infarcted heart for ischemic myocardial infarction[J]. Bioactive Materials, 2023, 27: 271-287. | IF: 16 Gao Y, Zhu Y, Wang H, et al. Lipid-mediated phase separation of AGO proteins on the ER controls nascent-peptide ubiquitination[J]. Molecular Cell, 2022, 82(7): 1313-1328. e8. | IF: 15.1 Chen X, Hao Y, Liu Y, et al. NAT10/ac4C/FOXP1 promotes malignant progression and facilitates immunosuppression by reprogramming glycolytic metabolism in cervical cancer[J]. Advanced Science, 2023, 10(32): 2302705. | IF: 12.8 Yang H H, Jiang H L, Tao J H, et al. Mitochondrial citrate accumulation drives alveolar epithelial cell necroptosis in lipopolysaccharide -induced acute lung injury[J]. Experimental & Molecular Medicine, 2022, 54(11): 2077-2091. |
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