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: cell specific or inducible.
Our SuRE™ technology does not deliver AI-based predictions that need to be verified by the customer but instead delivers the results of millions of actual wet-lab measurements in relevant (primary) cells and in vivo models.
To date Annogen has completed programs for companies like uniQure, Meira GTx and VectorY. In addition, we have generated elements in house that are available for licensing.
Besides promoters, the SuRE™ platform is also used to identify other regulatory elements supporting gene expression including introns, 5’UTRs, 3’UTRs as well as to test Capsids.
For more information on off-the-shelf products:
Identifying bespoke qualified human promoters driving cell specific or inducible
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.
Types of promoters we screen for:
Strong, stable and constitutive promoters
E.g. Housekeeping-like promoters to drive stable expression
Cell-type specific promoters
E.g. Neural promoters, blood-lineage specific promoters
Stimulation specific promoters
E.g. Neuroinflammation induced promoters, T-cell activation induced promoters
The program typically consists of 3 phases plus in vivo validation:
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
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).
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.