Applications in:
Promoter Screens For Cell & Gene Therapy
The efficacy and safety of cell and gene therapy strategies is in part determined by the correct expression profile of the therapeutic transgene. We perform bespoke SuRE™ screens to identify a human promoter with the optimal expression profile for your expression cassette, e.g. cell-specific or inducible.
Our SuRE™ technology does not deliver AI-based predictions that need to be verified by yourself, but rather delivers the results of millions of actual wet-lab measurements in relevant (primary) cells and/or in vivo models.
To date, Annogen has completed dozens of programs for leading biopharma including Orchard Therapeutics, uniQure, Meira GTx and VectorY. Our R&D team furthermore generates elements in-house, which are available for licensing.
Besides promoters, the SuRE™ platform can be used for the identification of other regulatory elements supporting gene expression including introns, 5’-UTRs, 3’-UTRs and capsid screening.
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For more information on off-the-shelf products:
Identifying bespoke qualified human promoters driving cell specific or inducible
transgene expression.
Cell and Gene Therapy strategies typically rely on stable overexpression of a therapeutic transgene or the Chimeric Antigen Receptor (CAR). In some cases, regulated expression is preferable, for example to achieve conditional expression of the therapeutic gene in situations like inflammation or an “armor” gene such as IL12. We apply our SuRE™ screening approach to identify the optimal (combination) of regulatory elements to drive transgene expression according to the customer’s needs.
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Examples of promoters we screen for:
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Strong, stable and constitutive promoters
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E.g. Housekeeping-like promoters to drive stable expression
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Cell-type specific promoters
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E.g. Neural promoters, blood-lineage specific promoters
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Stimulation specific promoters
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E.g. Neuroinflammation induced promoters, T-cell activation induced promoters
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The program typically consists of 3 phases plus in vivo validation:
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Phase 1 (~3 months)
Genome-wide analysis to find 200 stimulation or cell-type specific promoters and enhancers. Here we use genome-wide SuRE™ profiles in different cell types or stimulatory conditions to identify the most promising 200 promoters and enhancers for the next phase.
As we have a broad set of genome-wide profiles in an array of cell lines, many bespoke programs can start at phase 2
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Phase 2 (~3 months)
Hybrid screen: Analysis of all possible combinations between 200 promoters and enhancers (~160,000 combinations). Our experience is that by combining promoters and enhancers, expression levels as well as specificity can be greatly improved. So here we test al possible pairwise combinations between 200 elements (~160,000) which again are being tracked by at least 10 barcodes each to provide highly reliable data.
Phase 3 (~3 months)
Protein-level analysis of most promising hybrid promoter-enhancer combinations. Here we take the most promising 20 hybrids of the Phase 2 screen and now test them at the protein level (GFP expression in relevant cell types).
Phase 4
In vivo verification of top 5 hybrids. Typically, customers will now take up to 5 hybrids for in-house testing in vivo. However, we now also have the capability of providing in vivo work, depending on the exact analyses required.
