Genome Editing and Transcriptional Control System for Eukaryotic Cells

Request Number REQ3365116
Due Date February 9, 2018
Author Jos Cenens
Request for Proposal Details
RFP Title
Genome Editing and Transcriptional Control System for Eukaryotic Cells
RFP Description

NineSigma, representing a global health company, invites proposals for a system that can edit the genome of mammalian cells, particularly CHO-K1 or HEK293 derived cell lines. Ideally the system should be useable in both higher and lower eukaryotes (such as S. pombe, S. pastoris, S. cerevisiae). The desired system must be free of 3rd party background intellectual property and have the freedom to operate outside of the core technology patents in the genome editing and gene expression control space.


NineSigma’s client wants to engineer suitable host cell lines into novel phenotypes. This will involve gene knock-outs, insertions of genes of either inactive pathways or creating pathways that do not exist in the current host cell lines. Furthermore, they want to control gene transcription of endogenous and exogenously added genes (e.g., for synthetic biology or for generating induced pluripotent stem cells).  Of particular interest are CHO-K1 or HEK293-derived cell lines and the application of transcriptional control over genes in different eukaryotic cell types like S. pombe, S. pastoris, and S. cerevisiae.


The licensing terms for the commercial generation of new engineered cGMP-ready host cell lines using CRISPR-Cas9 system are expensive, so NineSigma’s client wants to identify an alternative system for editing the genome of eukaryotic cells. 


The proposed gene editing system needs convincing proof that the technology is viable and capable to meet the specifications.


Anticipated Project Phases or Project Plan 
The client will review submitted proposals and possibly ask clarifying questions before selecting the suitable candidates for collaboration. During the selection process, the client may execute non-disclosure agreements (NDA) with selected respondent(s), seek further information disclosure, and discuss specific development targets. The client will execute necessary agreement(s) with the selected respondent(s) and move to the commercial development phase.


Key Success Criteria

The successful technology will be capable to perform the functions associated with the CRISPR-Cas9 system:

  • Able to introduce double stranded DNA breaks to allow modifications to the genome
    • Have a specific endonuclease to cut double-stranded DNA at target site with sufficient activity (equal or better than Cas9 nuclease activity)
    • Possess the ability to separate double stranded DNA similarly to Cas9 helicase activity for the process to proceed efficiently
    • Endonuclease employed in the system is capable of being converted into a ‘nickase’ analogous to the CRISPR-Cas9n (e.g., Cas9_D10A mutation) system and can be used in an analogous way to the “double nickase” approach of CRISPR-Cas9n, creating a cut that has 5’ overhangs
    • The hindrance of the system to target DNA by nucleosomes/chromatin must be no worse than that observed in the CRISPR-Cas9 system
  • Allow facile target design, ideally without sequence constraints (e.g., no PAM sequence constraint)
  • Be compatible with other protein domains/modules (e.g., repressor, activator or chromatin modifying modules etc.) to allow transcriptional control of endogenous or exogenous genes.
  • Components of the system need to be easily expressed in microbial and mammalian cells (both for editing using transfected plasmids and to make the protein components of the targeting/nuclease complex)
  • Components can be easily purified and assembled in vitro so that the targeting complex can be transfected by a suitable method into mammalian cells (e.g., electroporation of Cas9n sgRNA targeting complex)


Possible Approaches

Possible approaches might include, but are not limited to:

  • Solutions derived from a different species of bacteria (e.g. not Streptococcus pyogenes, the species from where the CRISPR-Cas9 genome editing systems originates) or other relevant micro-organism. 
  • The system could be synthetic or designed as long as all are supported by data that meet the specifications
  • Partial solutions are of interest: client is prepared to look at a genome engineering based solution in the absence of transcriptional control solution and vice versa.



Approaches not of interest

The following approaches are not of interest:

  • Ideas without data to back up the proposal
  • Technologies with licensing terms that are too costly
  • Proposals from companies offering 3rd party licenses
  • Talen and Zinc finger based proposals



Preferred Collaboration types
  • Joint Development
  • Contract Research
  • Technology Licensing
  • Technology Acquisition
Items to be submitted

To respond, please make sure you are registered in NineSights, as it will prompt you to log in. Then, use the Response options at the top or bottom of the page. You may submit supplemental files in addition to your completed response form.


Appropriate responses will use the response form and address the following:

  • Non-confidential overview of proposed technology
  • Technical maturity of the approach (lab tested, ready to commercialize)
  • Data demonstrating that the approach can reach the targets
  • Obstacles to be overcome and remediation plans
  • Intellectual property situation and any constraints on freedom to operate
  • Brief overview of team behind the proposal and any information that prove your good understanding of genome engineering technology, its use in commercial environment, cGMP expertise


Other Request information includes:



Responses from companies (small to large), academic researchers, other research institutes, consultants, venture capitalists, entrepreneurs, or inventors are welcome. For example:


You represent a company that have demonstrated a proof of concept of a CRISPR alternative and are looking for a partner to commercialize it .

You represent a university research department that has promising results with a novel gene editing technology and is looking for financial support to finalize the development.



Area of Interest
Request Priority
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