FluidFM® CellEDIT Service


 Unique CRISPR cell line engineering service based on our FluidFM® technology.



Hard-to-transfect mammalian cells

TERT immortalized cells
Primary cells
iPSCs



Complex edits

Knock-in
Multiple knock-out
Large repair templates



Unmatched delivery speed

As fast as 6 weeks for fast growing cells


Our deliverables


  • 2 edited, independent monoclonal cell lines

  • 2 control cell lines

  • Final report incl. all data

Example customer projects

We do not shy away from editing primary cells, iPSCs, TERT-immortalized cell lines or other sensitive cell lines – adherent or in suspension. As we work vector-free, we also offer complex edits, like knock-in (KI) with large repair templates or multiple knock-outs (like triple KO). Benefit from novel disease models and complex edits thanks to FluidFM-based CellEDIT Service.



What was done
Cell type
Achieve­ment
Epithe­lial disease model
CRISPR KO
TERT-immortal­ized epithelial cells
First time ever suc­cess­ful CRISPR edit for this cell line
Undis­closed drug target
CRISPR KI
Undis­closed
5-10x higher HDR efficiency
Diabetes model
CRISPR KI
Hard-to-transfect cell line
CRISPR KI of sensitive cell
Skin model
CRISPR KOTERT-immortal­ized epi­der­mal cellsCRISPR KO of sensitive cell
Triple KO of CHO cells
CRISPR 3x KO in one step
CHO cells
3x CRISPR KO mono­clon­al cell line within 3 weeks

We combine the power of CRISPR with FluidFM® to provide engineered cell lines as a service 

Enabled by FluidFM® technology

Our proprietary FluidFM technology brings genome editing reagents directly into the nucleus. Safe, gentle to the cell, and vector-free! The basis of our technology is a force-controlled nanosyringe that allows precise injection of any soluble cargo directly into the cytoplasm or nucleus of any mammalian cell. And since we start with single cells, monoclonality of cell lines is guaranteed.

Image and Text
Monoclonality guaranteed

Because we start with a single cell, monoclonality of your cell lines is guaranteed.

FAQ

What is FluidFM CellEDIT?

FluidFM CellEDIT is a cell line engineering service by Cytosurge. We start with individually isolated single mammalian cells and inject a defined amount and concentration of editing reagents directly into the nucleus of these cells. Thanks to the high efficiency, only a few single cells must be edited and expanded into intrinsically monoclonal colonies.




Can you also perform edits on cells that are difficult to edit / hard-to-transfect?

In principle there are no limitations in editing cells with FluidFM technology. For certain cells though, some time is needed to find the proper culturing conditions to expand from a single cell. 


Can you do multiple edits at once?
  • Yes, we can do multiple edits at once. So far, we targeted three genes at once by using 3 different gRNA’s.

  • We do not target more that 3 sites at once to avoid potential DNA damage responses from the cells or unintended chromosomal rearrangements.

  • For more than 3 edits, we recommend performing multiple rounds of gRNA injections (Stack editing).

How fast can you deliver, what limits your editing speed?

We edit only single cells and therefore the speed is limited only by the natural doubling time of the particular cell type. Cell types unknown to us or never edited before, undergo an optimization procedure which adds in general roughly 3 weeks to the standard delivery time.

What influences the price for CellEDIT Services?

The price depends on:

  • Culturing conditions (e.g. co-culture, special media) have the biggest impact on the price

  • Type of edit: KI of sequences are more costly compared to simple KO as the editing efficiencies are lower and additional material is required. Further, multi-edits are more expensive than single edits for the same reasons

  • Extent of analysis 

  • Requested delivery time frame

  • Additional requests such as: more clones, etc.


What is the proforma quote?

The proforma quote is an automatically generated quote using your indications in the quote generator. We will check your request and send you a final quote within 1 working day. The final quote may be different from the proforma quote if special requests were included in the text fields of the quote generator.

How many edited clones do I get?

You will receive 2 edited clones and if needed, more edited clones can be selected in the quote generator.

How many reference clones do I get?

You will receive 2 monoclonal reference cells. The reference cell lines underwent the exact same culturing conditions as the edited cell lines. If needed, more reference clones can be selected in the quote generator.

How are the cells shipped?

The cells are shipped by courier on dry ice.
We are only shipping on Mondays and Tuesdays to avoid those cells are being stuck at customs over weekends.

What is the final report?

You will receive an extended final report including all the data obtained during the entire process e.g., high resolution gel pictures, which can be used for your publication.

Can you purchase a cell line for me?
  • Yes, if wished, we can purchase a cell line for you from established companies (e.g. Sigma, Thermo Fisher, ATCC). The costs for the cells, shipping and if required the licensing fees will be added to the quote.

  • For simplified licensing formalities and reduced costs, we recommend academic customers a) to send their cell lines, or b) to order cells in their name and ship them directly to Cytosurge.


Can I send my own cells?

Yes, this is possible. If you wish to send your own cells, please keep “User-supplied” in the quote generator selected.  With this, you also confirm that Cytosurge is allowed to perform edits on the indicated cell lines on your behalf.


Which shipping company should I use for sending the cells?
  • Cytosurge has partner companies that can take care of the shipping for you. 

  • Alternatively, you can use your preferred shipping company at your own risk. Please make sure that you inform Cytosurge ahead that your cells are being shipped and provide us with all required information (provider, tracking number).


Do I have to fill out customs documents if I ship cells?

We will provide all the required documentation for you. The cells are picked up by our courier service on Mondays or Tuesdays to avoid weekend shipping.

What culture media do you use?

We use your suggested media conditions or - in case you do not have preferences - use standard media conditions recommended by supplier and/or established cell culture brands (e.g. Gibco, Sigma, Takara, Stem Cell Technology)

How do you design the edits and identify the gRNA used?

  • In a first step, we import the genomic sequence of the target from ENSEMBL and annotate the coding exons. Usually, the first or second coding exon is used as template for identifying the gRNA binding site. In the case of multiple isoforms emanating from the gene, we will ask you to specify the isoforms to be targeted. Subsequently, we identify an exon that is common to all isoforms of interest and use it for gRNA design. 

  • Once we have identified the target site, we use two independent gRNA design tools: the Benchling Molecular Biology Suite gRNA design tool and the tool provided by IDT.

  • Both tools provide on and off-target scores for their suggested gRNA’s. The lists from both tools are compared and the gRNA found by both tools with the best scores is usually chosen. 

  • The finished design is audited by a 2nd scientist of the team and presented to you for approval before ordering reagents.


How do you design the ssDNA HDR templates for small insertions? (Point mutations or small inserts up to 120bp)
  • We obtain the genomic sequence from ENSEMBLE and annotate the site to be edited.

  • The target site is used as template for the HDR design tool from IDT.

  • The design provided by the tool is verified by one of our scientists for the following points:

    • gRNA location: must be as close as possible to the intended editing site.

    • Homology arms are between 30 and 75bp

    • Is the edited sequence still in frame?

    • Are the inserted/exchanged base pairs coding for the intended mutation?

  • The finished design is audited by a 2nd scientist of the team  and the reagents ordered


How do you design the HDR templates for large insertions? (More than 120bp)
  • We obtain the genomic sequence from ENSEMBLE and the sequence of the insert (e.g., GFP) and annotate the site to be edited.

  • The target site is used as template for the HDR design tool from IDT.

  • The design provided by the tool is verified by one of our scientists for the following points:

    • gRNA location: must be as close as possible to the intended editing site.

    • Homology arms are between 100 and 700bp

    • Is the edited sequence still in frame?

  • The finished design is audited by a 2nd scientist of the team and the reagents ordered


Where do you obtain your gRNA, Cas9 proteins, repair templates or primers from?
  • We order our reagents from IDT.

  • The exceptions are repair templates larger than 3000bp. These are bought from TWIST, Genwise or similar.


How do you analyze the cells before shipment?
  • Our standard analysis includes Sanger sequencing of targeted locus, alignment to the reference sequence, deconvolution of overlapping trace files using Indigo (Gear Genomics) and or DecodeR to determine the exact editing outcome. Routinely, we perform Mycoplasma and viability tests of cell lines before shipment.

  • If requested, we perform additional microbiology tests

  • Off-target evaluation: Sequencing of the 5 or 10 top off-target sites determined by the gRNA selection algorithm from IDT.  Whole genome sequencing is available upon request, additional quotation required).


What do you do with my generated cells after the CellEDIT service?

We keep backup vials of your generated cell lines, control cell lines and parent cell lines for a maximum of 3 months after shipping the order to you. The backup cells will be destroyed after this period. Upon request and a small fee, we are happy to store your cells for an extended period.

What is with IP rights of edited cells?

Cytosurge does not claim any IP on your cells or edits.

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