Custom Cell Lines

Horizon can tailor a custom cell line to your exact specifications

If you can't find your cell line of interest in our catalog, we can engineer a custom cell line for you. We use our cell line gene-editing platform to create a custom cell line to meet your exact specifications, whether that be a knockout, knockin, deletion, translocation or reporter cell line. Your custom cell line will be always delivered to you with its non-edited control.

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Use our custom cell lines to:

Horizon’s team has 10 years of experience in genome-editing using CRISPR, rAAV and Zinc Finger Nuclease (ZFNs) technologies which allows us to complete complex gene-editing projects in agreed timelines. If we haven’t engineered the cell line of your interest yet, our cell line characterization and optimization service will be included to maximize success of your gene editing project.

Custom Cell Line Generation

   

Starter

Standard

Premium

  • Sequence verified biallelic KO* or monoallelic KI via CRISPR-Cas9^#
  • DLD-1, HCT-116, or RKO cell line
  • Predesigned sgRNA (KO) or screening of 2 sgRNAs (KI)
  • Sequence verified biallelic KO* or monoallelic KI via CRISPR-Cas9^#
  • Any standard cancer cell line1**
  • Cell line characterization and optimization (if not used before)***
  • Predesigned sgRNA (KO) or screening of 2 sgRNAs (KI)
  • Mycoplasma Hoechst testing
  • Sequence verified biallelic KO* or monoallelic KI via any/all of our gene editing^ technologies
  • Any cell line1
  • Cell line characterization and optimization
  • Custom sgRNA design (KO) or screening of 5 sgRNAs (KI)
  • Mycoplasma Hoechst and culture isolation testing
  • Extended validation

From 10 weeks

From 13 weeks

From 20 weeks

*Knockouts by NHEJ-mediated frame-shift mutations. Defined deletions and gene disruption via targeted insertion are subject to knockin pricing. Some cell lines may be polyploid at some loci. Additional alleles may be modifed for an additional fee. **Possessed by Horizon or in ATCC for the commercial use. ***Characterization of more than one cell line is available for an additional fee. 1Cell line must tolerate single cell cloning and display adequate transfection efficiency. ^KI of additional alleles is available for an additional fee. #A plasmid donor carrying a selection cassette will be utilized.

Learn more about our gene-editing capabilities in induced pluripotent stem cells (iPSCs) and immune cells.

induced pluripotent stem cells (iPSC)     immune cells

Case studies

Case study I: CRISPR-Mediated Homozygous Knockout
Purpose: Disruption of the gene X in the human melanoma A375 cell line (copy number = 3)

Workflow

  • Transfection with plasmids expressing Cas9 nuclease and a validated gRNA
  • Derivation of single cell clones
  • Screening of the clones
  • Genotype analysis by Sanger sequencing

Targeting Strategy


 

Targeting

  • Sequencing of individual alleles of 7 clones (via molecular cloning) demonstrated that 3 clones had out of frame deletions on all 3 alleles
  • Sequencing of transcripts from individual alleles indicated out of frame deletions were also present at the RNA level for two of the three clones

Clone 1

Case Study II: Heterozygous Knockin in Leukemia Cells 
Purpose: To create a heterozygous point mutation K700E (AAA -> GAA) in SF3B1 in chronic myelogenous leukemia (CML) K562 cells using rAAV

Workflow

  • Design of rAAV targeting vectors
  • rAAV infection
  • Plating of infected clones
  • PCR and Sanger sequencing to identify KI pools
  • Derivation of single cell clones
  • Screening of the clones by PCR
  • Genotype analysis by Sanger sequencing

Vector Design

 

Targeting

Targeted allele-specific sequencing shows that both clones contain the K700E mutation. Non-allele specific sequencing shows a ratio of 50:50 of the mutant to wild-type peaks, as would be expected for a 2N locus.

 
What else can Horizon do to support your project?

Cell Line Characterization



  • Whole genome sequencing
  • Screening for off-target effects
  • mRNA expression analysis
  • Protein analysis

Functional Validation



Other Assay Services 



In Vivo Studies



Request more information about our assays

Related materials

Watch our webinar on gene editing Click here
Read our blog post on improving gene-editing success with optimizing cell line screening procedure Click here
Learn how panels of engineered cell lines can be used for predicting patient drug responses Click here
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