Cell Line Engineering Service

Comprehensive gene regulation cell line construction services

Gene Editing Technologies

The CRISPR/Cas9 system uses guide RNA (sgRNA) to direct the Cas9 nuclease to target specific genomic loci, generating DNA double-strand breaks that are then repaired through cellular DNA repair mechanisms — non-homologous end joining (NHEJ) or homology-directed repair (HDR) — enabling precise editing of target genes. Building upon this foundation, multiple derivative technologies have further expanded the application boundaries of gene editing:

Transcriptional Regulation Systems (CRISPRa/CRISPRi) Fusing catalytically dead Cas9 (dCas9) with transcriptional activation domains (e.g., VP64, p65) or transcriptional repression domains (e.g., KRAB), and targeting specific gene promoter regions under sgRNA guidance, enables reversible upregulation or downregulation of target gene expression without altering the genomic sequence itself.

Epigenome Editing Systems Fusing dCas9 with epigenetic modifier enzymes such as DNMT3A or TET1 enables directed rewriting of DNA methylation or demethylation status at specific genomic regions, with heritable modification effects that persist through cell division.

Base Editors By fusing cytosine or adenine deaminases with Cas9 nickase, precise single-base conversions are achieved without generating DNA double-strand breaks.

Prime Editors By fusing reverse transcriptase with Cas9 nickase and combining with engineered pegRNA, precise replacement, insertion, or deletion of target sequences is achieved.

Gene-Edited Cell Lines & Applications

Gene-edited cell lines are fundamental tools for functional genomics research. By introducing predefined genetic modifications into specific cell lines, researchers can dissect gene functions, screen drug targets, construct disease models, and evaluate the feasibility of gene therapy strategies under controlled genetic backgrounds.

Gene Function Research　 Systematically dissect gene biological functions in specific cellular contexts, establishing genotype-phenotype causal associations.
Drug Target Discovery & Validation　 Construct target validation cell models via gene knockout or regulation, evaluating the biological effects of target intervention.
Disease Model Construction　 Introduce pathogenic mutations or key gene modifications to construct cell models for diseases such as cancer and genetic disorders.
Gene Therapy Strategy Evaluation　 Evaluate the efficacy of gene editing or gene therapy strategies at the cellular level.
Synthetic Lethality Target Screening　 Construct cell lines with specific gene deficiencies, providing experimental systems for synthetic lethality drug screening.

Gene Regulation Types & Technical Services

Regulation Type	Technical Route	Delivery Standard	Delivery Cycle (Est.)
Gene Knockout (KO)	CRISPR/Cas9 plasmid / lentivirus / RNP transduction	Monoclonal, Sanger sequencing confirmed frameshift / fragment deletion	8-14 weeks
Endogenous Activation (CRISPRa)	dCas9-SAM system lentiviral transduction	Pooled clone, qPCR confirmed target gene transcriptional upregulation	8-12 weeks
Endogenous Inhibition (CRISPRi)	dCas9-KRAB system lentiviral transduction	Pooled clone, qPCR confirmed target gene transcriptional downregulation	8-12 weeks
Site-Directed Mutagenesis	HDR template + CRISPR knock-in strategy	Monoclonal, Sanger sequencing confirmed mutant sequence	12-20 weeks
Site-Directed Methylation	dCas9-DNMT3A fusion system lentiviral transduction	Pooled clone, bisulfite sequencing confirmed methylation status	8-14 weeks
Site-Directed Demethylation	dCas9-TET1 fusion system lentiviral transduction	Pooled clone, bisulfite测序 confirmed demethylation status	8-14 weeks
Conventional Overexpression	Overexpression vector lentiviral transduction	Pooled clone, qPCR confirmed mRNA overexpression	6-10 weeks
Conventional Knockdown	shRNA lentiviral transduction	Pooled clone, qPCR confirmed target mRNA knockdown level	6-10 weeks

Workflow

Project Evaluation

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Vector Design & Construction

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Cell Transduction

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Positive Cell Enrichment / Monoclonal Screening

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Gene Editing Event Validation

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Expansion & Quality Control

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Delivery

Technical route feasibility depends on gene locus and cell type. Specific protocols require technical consultation to confirm.