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Point Mutations Cell Lines
Engineer disease-specific cell lines with Cyagen's targeted gene editing technology, delivering validated point mutations in just 6 weeks. Achieve up to 87% HDR efficiency across challenging cell types, backed by comprehensive QC documentation and guaranteed homozygous clones.
Comprehensive Deliverables
Receive homozygous clones with detailed QC reports and biweekly updates.
Flash Turnaround
Obtain homozygous clones in as fast as 6 weeks.
Optimized α-Donor System
Achieves HDR efficiency up to 87% for precise point mutations.
Overview
Workflow
FAQs
Overview
Point Mutation Cell Line: Precision Meets Efficiency
Cyagen's targeted gene editing technology delivers precise genomic modifications with unprecedented efficiency. Our validated protocol achieves 87% HDR rates and 70% homozygous outcomes across diverse cell lines, enabling rapid disease modeling and drug discovery with comprehensive quality validation.
Explore Ready-to-Use Mouse Models
Discover over 18,000 validated mouse strains—including knockout, conditional knockout, and humanized models—covering 20+ research areas such as oncology, neurology, and metabolism. All models are supported by detailed genotype data and guaranteed quality, helping you fast-track discovery with confidence.
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Browse validated human KO cell lines by gene or cell type
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Access characterized human tumor lines by cancer type or genetic status.
Targeted Gene Editing
AI-enhanced HDR technology for precise in vivo genome engineering
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Search and order AAVs by serotype, gene, or function
MouseAtlas Model Library
Search and access curated genetically engineered mouse strains
Cell Immunotherapy CRO Platform
CAR-T/NK development with custom vectors, models, and in vivo validation
Workflow
Workflow and Delivery
Our cell line development services provide a comprehensive and streamlined workflow for generating genetically modified cell lines, ensuring quality, reproducibility, and fast delivery. Below, you will find detailed descriptions of the workflows, estimated timelines, and deliverables for each type of cell line, tailored to your research needs.
Point Mutation Cell Lines
iPSC Point Mutation Cell Lines
Point Mutation Cell Lines
Point Mutation Cell Line Development & Delivery
Cyagen’s Point Mutation Cell Line development workflow ensures precise, efficient gene editing. From initial design to final QC, we offer a transparent, quality-controlled process to deliver reliable point mutation cell lines within the expected timeframe.
Deliverables & QC
Cell Type Example Cell Lines Deliverables Quality Control (QC)
Tumor & Immune Cells Jurkat, HepG2, SK-MES-1 Point mutation clones + matched WT controls (2 vials each, 10⁶ cells/vial) + validation report PCR, Sanger sequencing
Non-Cancerous Cells HK-2, AC16
Workflow & Timeline
Stage Description Turnaround Time
Cell Expansion & QC 1. Mycoplasma and sterility testing.
2. Cell proliferation assessment.
3. Primer design for sequencing the target region.
 1-2 weeks
Guide Molecule & Donor Vector Design 1. Design and synthesize guide molecules for gene targeting.
2. Construct donor template.
 2-5 weeks
Electroporation & Clone Selection 1. Electroporate guide molecules into target cells.
2. Screen single clones, extract genomic DNA, and validate point mutation through PCR and sequencing.
 4-6 weeks
Cryopreservation & Final QC 1. Sequence analysis of the mutation site.
2. Mycoplasma and sterility testing.
3. Viability testing.
 1-2 weeks
Note:
  • Total Estimated Time: 8-15 weeks
  • Time may vary depending on cell type and project complexity.
iPSC Point Mutation Cell Lines
iPSC Point Mutation Cell Line Development & Delivery
Our iPSC Point Mutation Cell Line development process is optimized for high-quality gene editing while maintaining the pluripotency of the cells. With a dedicated workflow and expert oversight, we ensure robust, validated iPSC lines that meet the rigorous standards of your research.
Deliverables & QC
Cell Type Example Cell Lines Deliverables Quality Control (QC)
iPSC Point Mutation Cell Lines H1, H9, iPSC Differentiated or undifferentiated cell lines PCR, Sanger sequencing, Immunofluorescence (IF)
Workflow & Timeline
Stage Description Turnaround Time
Cell Expansion & QC 1. Mycoplasma and sterility testing.
2. Cell proliferation assessment.
3. Primer design for sequencing the target region.
 1-2 weeks
Guide Molecule & Donor Vector Design 1. Design and synthesize guide molecules for gene targeting.
2. Construct donor template.
 2-5 weeks
Electroporation & Clone Selection 1. Electroporate guide molecules into target iPSCs.
2. Screen single clones, extract genomic DNA, and validate point mutation through PCR and sequencing.
 4-6 weeks
Cryopreservation & Final QC 1. Sequence analysis of the mutation site.
2. Mycoplasma and sterility testing.
3. Viability testing.
4. Immunofluorescence staining to verify pluripotency markers.
 2 weeks
Note:
  • Total Estimated Time: 8-12 weeks
  • Turnaround time may vary for specialized iPSC lines.
FAQs
Frequently Asked Questions (FAQs)
What's your approach for introducing silent mutations in PAM sites when engineering temperature-sensitive variants? How do you validate the temperature-dependent phenotype?
We design silent mutations using advanced computational tools to disrupt PAM sites without altering amino acid sequences. Temperature sensitivity validation involves phenotype assays measuring growth rate or protein activity at permissive versus restrictive temperatures, complemented by functional sequencing. For example, temperature-sensitive BRAF mutants would be assessed for MAPK pathway activity via p-ERK Western blot under varying temperature conditions.
For BRAF V600E mutation in melanoma cell lines, how do you verify the activation status of the downstream MAPK pathway post-editing? Do you include Western blot validation?
We confirm BRAF V600E functionality and MAPK pathway activation through Western blot analysis of phosphorylated ERK (p-ERK), a key pathway marker. Additionally, we use VE1 antibody staining to validate the BRAF V600E mutation specifically. These validations ensure the edited cells exhibit expected pathway activation, which is critical for modeling therapeutic responses accurately.
What's your protocol for introducing multiple point mutations within the same exon of TP53? Can you achieve this without affecting the surrounding regulatory elements?
Our approach combines multiplex gRNA design with our proprietary α-donor system to introduce multiple mutations within a single exon. We utilize homology-directed repair (HDR) to ensure precision and avoid disruption of adjacent regulatory elements. Post-editing confirmation includes Sanger sequencing and functional assays, such as CypD interaction checks for TP53 exon-6 truncations, to maintain genomic integrity.
For iPSC lines, what's your strategy to maintain pluripotency markers (OCT4, NANOG) during the gene editing process? Do you provide flow cytometry data for stemness verification?
We prioritize pluripotency maintenance by optimizing the delivery of gene modification components to minimize cellular stress, and employing footprint-free editing through our proprietary α-donor system. Post-editing validation includes immunofluorescence staining for OCT4, NANOG, and SOX2. While our standard reports include immunofluorescence images, we can provide quantitative flow cytometry data for stemness verification upon request.
Can Cyagen achieve homozygous KRAS G12D mutations in HCT116 cells while maintaining their microsatellite instability (MSI) status? What's your validation method for MSI verification post-editing?
Yes, our Cell Gene Editing System with targeted gene editing technology enables precise homozygous KRAS G12D mutations in HCT116 cells while preserving MSI status. We validate MSI stability using PCR-capillary electrophoresis to analyze five microsatellite loci (including BAT25 and BAT26), following standardized colorectal cancer research protocols. This ensures no unintended genomic disruptions occur during the editing process.
What Customers Say About Cyagen
Violet Shimmer
Stanford University
The service provided to us by Cyagen is now in press at Nature as an article.
Scarlett Rouge
Seattle Children’s Hospital
We are very pleased with the state-of-the-art professional transgenic services provided by Cyagen for our study published recently in Nature. We continue to use Cyagen’s transgenic services as it allows us to do better and more efficient research with transgenic mice.
Request a Custom Cell Line Consultation
Tell us about your cell line project needs. Our specialists are ready to support your research with tailored solutions.
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