Design of a synthesis‐friendly hypoxia‐responsive promoter for cell‐based therapeutics
Abstract Towards the goal of making ‘smart’ cell therapies, one that recognizes disease conditions (e.g. hypoxia) and then produces mitigating biologics, it is important to develop suitable promoters. Currently, hypoxia responsive promoters are composed of strongly repeated sequences containing hypo...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Wiley-VCH
2021-12-01
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| Series: | Engineering in Life Sciences |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/elsc.202100045 |
| _version_ | 1818399207600422912 |
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| author | Chee Ka Candice Lam Kevin Truong |
| author_facet | Chee Ka Candice Lam Kevin Truong |
| author_sort | Chee Ka Candice Lam |
| collection | DOAJ |
| description | Abstract Towards the goal of making ‘smart’ cell therapies, one that recognizes disease conditions (e.g. hypoxia) and then produces mitigating biologics, it is important to develop suitable promoters. Currently, hypoxia responsive promoters are composed of strongly repeated sequences containing hypoxia response elements upstream of a minimal core promoter. Unfortunately, such repeated sequences have inherent genomic instability that may compromise the long‐term consistency of cell‐based therapeutics. Thus, we designed a synthesis‐friendly hypoxia‐inducible promoter (named SFHp) that has GC content between 25% and 75% and no repeats greater than 9 base pairs. In HEK293 cells stably integrated with genes regulated by synthetic SFHp, we demonstrated inducible reporter expression with fluorescent proteins, cell morphology rewiring with our previously engineered RhoA protein and intercellular cell signalling with secreted cytokines. These experiments exemplify the potential usage of SFHp in cell‐based therapeutics with integrated genetic circuits that inducibly respond to the disease microenvironment. |
| first_indexed | 2024-12-14T07:17:01Z |
| format | Article |
| id | doaj.art-24ee11923f6240e090509d3fa57cb09b |
| institution | Directory Open Access Journal |
| issn | 1618-0240 1618-2863 |
| language | English |
| last_indexed | 2024-12-14T07:17:01Z |
| publishDate | 2021-12-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | Engineering in Life Sciences |
| spelling | doaj.art-24ee11923f6240e090509d3fa57cb09b2022-12-21T23:11:42ZengWiley-VCHEngineering in Life Sciences1618-02401618-28632021-12-01211284885610.1002/elsc.202100045Design of a synthesis‐friendly hypoxia‐responsive promoter for cell‐based therapeuticsChee Ka Candice Lam0Kevin Truong1Institute of Biomedical Engineering University of Toronto Toronto ON CanadaInstitute of Biomedical Engineering University of Toronto Toronto ON CanadaAbstract Towards the goal of making ‘smart’ cell therapies, one that recognizes disease conditions (e.g. hypoxia) and then produces mitigating biologics, it is important to develop suitable promoters. Currently, hypoxia responsive promoters are composed of strongly repeated sequences containing hypoxia response elements upstream of a minimal core promoter. Unfortunately, such repeated sequences have inherent genomic instability that may compromise the long‐term consistency of cell‐based therapeutics. Thus, we designed a synthesis‐friendly hypoxia‐inducible promoter (named SFHp) that has GC content between 25% and 75% and no repeats greater than 9 base pairs. In HEK293 cells stably integrated with genes regulated by synthetic SFHp, we demonstrated inducible reporter expression with fluorescent proteins, cell morphology rewiring with our previously engineered RhoA protein and intercellular cell signalling with secreted cytokines. These experiments exemplify the potential usage of SFHp in cell‐based therapeutics with integrated genetic circuits that inducibly respond to the disease microenvironment.https://doi.org/10.1002/elsc.202100045biosensorhypoxiainducible expressionpromoter designsynthetic promoters |
| spellingShingle | Chee Ka Candice Lam Kevin Truong Design of a synthesis‐friendly hypoxia‐responsive promoter for cell‐based therapeutics Engineering in Life Sciences biosensor hypoxia inducible expression promoter design synthetic promoters |
| title | Design of a synthesis‐friendly hypoxia‐responsive promoter for cell‐based therapeutics |
| title_full | Design of a synthesis‐friendly hypoxia‐responsive promoter for cell‐based therapeutics |
| title_fullStr | Design of a synthesis‐friendly hypoxia‐responsive promoter for cell‐based therapeutics |
| title_full_unstemmed | Design of a synthesis‐friendly hypoxia‐responsive promoter for cell‐based therapeutics |
| title_short | Design of a synthesis‐friendly hypoxia‐responsive promoter for cell‐based therapeutics |
| title_sort | design of a synthesis friendly hypoxia responsive promoter for cell based therapeutics |
| topic | biosensor hypoxia inducible expression promoter design synthetic promoters |
| url | https://doi.org/10.1002/elsc.202100045 |
| work_keys_str_mv | AT cheekacandicelam designofasynthesisfriendlyhypoxiaresponsivepromoterforcellbasedtherapeutics AT kevintruong designofasynthesisfriendlyhypoxiaresponsivepromoterforcellbasedtherapeutics |