A genetic safeguard for eliminating target genes in synthetic probiotics in a gut environment
Summary: To meet the emerging demand for constraining engineered probiotic activities, many biocontainment studies explore strategies that involve killing engineered microbes, which often create basal levels of cytotoxicity that hamper cell fitness and performance. Here, we explored a circuit design...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-08-01
|
| Series: | iScience |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S258900422501288X |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1839631224357781504 |
|---|---|
| author | Nhu Nguyen Miaomiao Wang Lin Li Clement T.Y. Chan |
| author_facet | Nhu Nguyen Miaomiao Wang Lin Li Clement T.Y. Chan |
| author_sort | Nhu Nguyen |
| collection | DOAJ |
| description | Summary: To meet the emerging demand for constraining engineered probiotic activities, many biocontainment studies explore strategies that involve killing engineered microbes, which often create basal levels of cytotoxicity that hamper cell fitness and performance. Here, we explored a circuit design that destroys the engineered genetic materials in a probiotic strain, instead of killing these cells, under non-permissive conditions. Our safeguard circuit involves a two-layered transcriptional regulatory circuit to control the expression of a CRISPR system that targets the engineered genes for degradation. In Escherichia coli Nissle 1917 (EcN), the biocontainment system did not hamper cell fitness, and it continuously scavenged and destroyed the target, promoting complete elimination of engineered genetic materials and activities. We demonstrated that the engineered probiotics maintained its activities for 7 days in a mouse model when the permissive signal was supplied constantly, but the activities became undetectable within two days upon the cease of signal supply. |
| format | Article |
| id | doaj-art-a8364054e76d463f94f22e3ece4ed070 |
| institution | Matheson Library |
| issn | 2589-0042 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj-art-a8364054e76d463f94f22e3ece4ed0702025-07-12T04:46:33ZengElsevieriScience2589-00422025-08-01288113027A genetic safeguard for eliminating target genes in synthetic probiotics in a gut environmentNhu Nguyen0Miaomiao Wang1Lin Li2Clement T.Y. Chan3Department of Biomedical Engineering, University of North Texas, Denton, TX, USADepartment of Biomedical Engineering, University of North Texas, Denton, TX, USADepartment of Biomedical Engineering, University of North Texas, Denton, TX, USADepartment of Biomedical Engineering, University of North Texas, Denton, TX, USA; BioDiscovery Institute, University of North Texas, Denton, TX, USA; Corresponding authorSummary: To meet the emerging demand for constraining engineered probiotic activities, many biocontainment studies explore strategies that involve killing engineered microbes, which often create basal levels of cytotoxicity that hamper cell fitness and performance. Here, we explored a circuit design that destroys the engineered genetic materials in a probiotic strain, instead of killing these cells, under non-permissive conditions. Our safeguard circuit involves a two-layered transcriptional regulatory circuit to control the expression of a CRISPR system that targets the engineered genes for degradation. In Escherichia coli Nissle 1917 (EcN), the biocontainment system did not hamper cell fitness, and it continuously scavenged and destroyed the target, promoting complete elimination of engineered genetic materials and activities. We demonstrated that the engineered probiotics maintained its activities for 7 days in a mouse model when the permissive signal was supplied constantly, but the activities became undetectable within two days upon the cease of signal supply.http://www.sciencedirect.com/science/article/pii/S258900422501288XTechniques in geneticsApplied microbiologyMicrobial geneticsBiomolecular engineering |
| spellingShingle | Nhu Nguyen Miaomiao Wang Lin Li Clement T.Y. Chan A genetic safeguard for eliminating target genes in synthetic probiotics in a gut environment iScience Techniques in genetics Applied microbiology Microbial genetics Biomolecular engineering |
| title | A genetic safeguard for eliminating target genes in synthetic probiotics in a gut environment |
| title_full | A genetic safeguard for eliminating target genes in synthetic probiotics in a gut environment |
| title_fullStr | A genetic safeguard for eliminating target genes in synthetic probiotics in a gut environment |
| title_full_unstemmed | A genetic safeguard for eliminating target genes in synthetic probiotics in a gut environment |
| title_short | A genetic safeguard for eliminating target genes in synthetic probiotics in a gut environment |
| title_sort | genetic safeguard for eliminating target genes in synthetic probiotics in a gut environment |
| topic | Techniques in genetics Applied microbiology Microbial genetics Biomolecular engineering |
| url | http://www.sciencedirect.com/science/article/pii/S258900422501288X |
| work_keys_str_mv | AT nhunguyen ageneticsafeguardforeliminatingtargetgenesinsyntheticprobioticsinagutenvironment AT miaomiaowang ageneticsafeguardforeliminatingtargetgenesinsyntheticprobioticsinagutenvironment AT linli ageneticsafeguardforeliminatingtargetgenesinsyntheticprobioticsinagutenvironment AT clementtychan ageneticsafeguardforeliminatingtargetgenesinsyntheticprobioticsinagutenvironment AT nhunguyen geneticsafeguardforeliminatingtargetgenesinsyntheticprobioticsinagutenvironment AT miaomiaowang geneticsafeguardforeliminatingtargetgenesinsyntheticprobioticsinagutenvironment AT linli geneticsafeguardforeliminatingtargetgenesinsyntheticprobioticsinagutenvironment AT clementtychan geneticsafeguardforeliminatingtargetgenesinsyntheticprobioticsinagutenvironment |