Knockin Cell Lines
Drive your experiments using site-specific modifications that support gene regulation studies, disease modeling, and drug screening. Our knockin cell lines offer consistent insertion of reporters, point mutations, or fusion tags, delivering validated solutions for actionable insights in your next project.
Rapid Turnaround
Obtain your customized knockout cell line in as little as 2 weeks.
Expert-Guided
Free tech consultation on model selection and experimental alignment in minute
Guaranteed Outcomes
We deliver as promised—or your money back.
Overview
Workflow
FAQs
Overview
Knockin Cell Lines: Defined Genomic Modifications
Cyagen's targeted gene editing cell lines feature documented site-specific DNA insertions at predetermined loci. Each model undergoes rigorous validation with full QC documentation, enabling researchers to study gene function, track proteins, and model diseases with high experimental reproducibility.
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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Workflow
Workflow and Delivery
Cyagen offers a streamlined process for generating Knockin (KI) cell
lines, ensuring precise genetic modifications for various research
applications. Our workflow covers all stages, from gene design to final
cell line validation, with strict quality control at each step. We
deliver well-characterized, stable cell lines along with comprehensive
reports and sequencing data, providing reliable models for your
research.
Knockin Cell Lines
iPSC Knockin Cell Lines
Knockin Cell Lines
Knockin Cell Line Development & Delivery
Cyagen provides a streamlined process for Knockin
(KI) cell line development, ensuring accurate gene
insertion and high-quality models for various
research applications. The following workflow
outlines the typical steps involved in generating
Knockin cell lines, with estimated turnaround time
and deliverables.
Deliverables & QC
| Cell Type | Example Cell Lines | Deliverables | Quality Control (QC) |
|---|---|---|---|
| Tumor & Immune Cells | Jurkat, HepG2, SK-MES-1 | Knockin 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. Donor vector design and construction |
5-7 weeks |
| Electroporation & Clone Selection |
1. Electroporate guide molecules
into target cells. 2. Screen single clones, extract genomic DNA, and validate the knockin via PCR and sequencing. |
4-8 weeks |
| Cell Cryopreservation & QC |
1. Sequence analysis of the knockin
site. 2. Mycoplasma and sterility testing. 3. Viability testing. |
1-2 weeks |
Note:Total Estimated Time:
11-19 weeks
iPSC Knockin Cell Lines
iPSC Knockin Cell Line Development & Delivery
For iPSC Knockin Cell Line development, Cyagen
offers an optimized workflow to ensure precise gene
insertion while preserving the pluripotency and
differentiation potential of the stem cells. Below
is the workflow with estimated timelines and
deliverables.
Deliverables & QC
| Cell Type | Example Cell Lines | Deliverables | Quality Control (QC) |
|---|---|---|---|
| iPSC Knockin 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. Donor vector design and construction |
5-6 weeks |
| Electroporation & Clone Selection |
1. Electroporate guide molecules
into target iPSCs. 2. Screen single clones, extract genomic DNA, and validate the knockin via PCR and sequencing. |
4-8 weeks |
| Cell Cryopreservation & QC |
1. Sequence analysis of the knockin
site. 2. Mycoplasma and sterility testing. 3. Viability testing. 4. Immunofluorescence staining to verify pluripotency markers. |
1-2 weeks |
Note:Total Estimated Time:
11-18 weeks
FAQs
Frequently Asked Questions (FAQs)
For APOE4 point mutation knockins for Alzheimer's research, can you provide functional validation that the mutation affects protein folding?
We deliver fully sequenced and clonally validated APOE4-mutated cell lines, including protein expression level checks. While functional validation (e.g., folding assays) is not standard, we collaborate with clients to design tailored experiments or recommend downstream partners for phenotypic analysis. Our HepG2 case study highlights our ability to support functional research pipelines.
What specific validation methods do you use to confirm the absence of off-target effects beyond the primary insertion site?
We employ:
●Sanger sequencing to verify target locus integrity.
●PCR-based screening to confirm insert size and orientation.
●Cell viability assays to rule out unintended functional impacts.
While whole-genome off-target analysis is not explicitly mentioned, our stringent QC (100% contamination-free cells and dual mycoplasma/bacterial testing) ensures reliable outcomes.
●Sanger sequencing to verify target locus integrity.
●PCR-based screening to confirm insert size and orientation.
●Cell viability assays to rule out unintended functional impacts.
While whole-genome off-target analysis is not explicitly mentioned, our stringent QC (100% contamination-free cells and dual mycoplasma/bacterial testing) ensures reliable outcomes.
Do you offer humanized protein expression in mouse cell lines for antibody development projects?
Yes. Our KI services include humanized protein expression in mouse cell lines, enabling applications like antibody development. For example, we generated SR-BI mutant HepG2 cells to study humanized protein interactions in mice. Custom requests for species-specific protein swaps or epitope tagging are available.
Can you perform multiple gene edits simultaneously in the same cell line, and how does this affect validation timelines?
Yes, we support simultaneous multi-gene edits, as demonstrated in our case study delivering a double-point mutation HepG2 cell line. Validation timelines may extend slightly due to additional sequencing and quality checks, but our standardized workflow (including single-cell cloning and multi-step sequencing) ensures thorough characterization. Timelines are project-specific and discussed during quoting.
What is your typical efficiency rate for knockin of large inserts (>2kb) in hard-to-transfect cell lines like primary neurons?
Our Cell iGeneEditor™ system achieves up to 90% editing efficiency for gene knockins (KIs). While insert size and cell type influence outcomes, our optimized homologous recombination protocols and extensive experience with diverse cell types (including neuronal lines like SH-SY5Y and U251) ensure robust results. For hard-to-transfect cells, we conduct pre-experiments to assess feasibility and refine delivery methods. Primary neurons are not explicitly listed in our cell bank, but contact us for custom project evaluations.
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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