Difference between revisions of "Part:BBa K4375018"

Latest revision as of 04:52, 11 October 2022

MiniNano for bacterial surface display

This construction consists of 4 described parts. These are: Codon optimalised INPNC for Surface Display (BBa_K4375007), flexible GS-linker (BBa_K4375010), miRFP670 nano (BBa_K4375008), J23101* (BBa_K4375003). They allow the bacterium to be expelled to the tumor surface, thus allowing its visualisation and colonisation on the tumour surface.

Usage and Biology

The MiniNano is a compact construct meant for extracellular protein (like tumor-specific receptor, antigen, etc.) detection via fluorescence. This protein has 3 domains. (1) INPNC surface display module (INPNC codes for N- and C- terminal domain of Ice Nucleation Protein (INP) from Pseudomonas syringe, and it is used for displaying proteins on bacteria's outer membrane (BBa_K4375007). It is linked to (by a flexible GS-linker, BBa_K4375010) (2) a small, near-infrared emitting fluorescent protein, miRFP670 nano (BBa_K4375008), which is connected to an anti-EGFR specific Nanobody, 7D12, via llama-derived IgA hinge region linker (BBa_K4375011). Moreover, it contains a Myc epitope tag, so the specific interaction can be detected with labelled antibodies. This complex detection device is expressed under the constitutive promoter J23101* (BBa_K4375003) The Nanobody in the construct can be easily swapped with other protein domains, as its sequence is framed by BamHI and SphI restriction sites

Figure 1: MiniNano composite scheme.

Sequence and Features

References

Wu, P.-H.; Giridhar, R.; Wu, W.-T. Surface Display of Transglucosidase onEscherichia Coli by Using the Ice Nucleation Protein ofXanthomonas Campestris and Its Application in Glucosylation of Hydroquinone. Biotechnology and Bioengineering 2006, 95 (6), 1138–1147. https://doi.org/10.1002/bit.21076.

https://2012.igem.org/Team:Penn/Team

„Myc-tag: An epitope tag for protein characterization, protein interaction analysis, and purification.” https://www.ptglab.com/news/blog/myc-tag-an-epitope-tag-for-protein-characterization-protein-interaction-analysis-and-purification/

Li, D.; Ren, J.; Ji, F.; Peng, Q.; Teng, H.; Jia, L. Peptide Linker Affecting the Activity Retention Rate of VHH in Immunosorbents. Biomolecules 2020, 10 (12), 1610. https://doi.org/10.3390/biom10121610

Karl R. Schmitz, Atrish Bagchi, Rob C. Roovers, Paul M.P. van Bergen en Henegouwen, Kathryn M. Ferguson, Structural Evaluation of EGFR Inhibition Mechanisms for Nanobodies/VHH Domains, Structure, Volume 21, Issue 7, 2013, Pages 1214-1224, ISSN 0969-2126, https://doi.org/10.1016/j.str.2013.05.008.

Lwin, T. M.; Turner, M. A.; Nishino, H.; Amirfakhri, S.; Hernot, S.; Hoffman, R. M.; Bouvet, M. Fluorescent Anti-CEA Nanobody for Rapid Tumor-Targeting and Imaging in Mouse Models of Pancreatic Cancer. Biomolecules 2022, 12 (5), 711. https://doi.org/10.3390/biom12050711.

De Meyer, T.; Muyldermans, S.; Depicker, A. Nanobody-Based Products as Research and Diagnostic Tools. Trends in Biotechnology 2014, 32 (5), 263–270. https://doi.org/10.1016/j.tibtech.2014.03.001.

Turner, M. A.; Lwin, T. M.; Amirfakhri, S.; Nishino, H.; Hoffman, R. M.; Yazaki, P. J.; Bouvet, M. The Use of Fluorescent Anti-CEA Antibodies to Label, Resect and Treat Cancers: A Review. Biomolecules 2021, 11 (12), 1819. https://doi.org/10.3390/biom11121819.

Lee, J. H.; Lee, S.-W. The Roles of Carcinoembryonic Antigen in Liver Metastasis and Therapeutic Approaches. Gastroenterology Research and Practice 2017, 2017, 1–11. https://doi.org/10.1155/2017/7521987.

Grunnet, M.; Sorensen, J. B. Carcinoembryonic Antigen (CEA) as Tumor Marker in Lung Cancer. Lung Cancer 2012, 76 (2), 138–143. https://doi.org/10.1016/j.lungcan.2011.11.012.

Boon L., Yutong Y.; Reprogramming Synthetic Cells for Targeted Cancer Therapy 2022; 11 (3), 1349-1360 DOI: 10.1021/acssynbio.1c00631

Oliinyk, O. S.; Shemetov, A. A.; Pletnev, S.; Shcherbakova, D. M.; Verkhusha, V. V. Smallest Near-Infrared Fluorescent Protein Evolved from Cyanobacteriochrome as Versatile Tag for Spectral Multiplexing. Nature Communications 2019, 10 (1). https://doi.org/10.1038/s41467-018-08050-8.

Oliinyk, O.; Chernov, K.; Verkhusha, V. Bacterial Phytochromes, Cyanobacteriochromes and Allophycocyanins as a Source of Near-Infrared Fluorescent Probes. International Journal of Molecular Sciences 2017, 18 (8), 1691. https://doi.org/10.3390/ijms18081691.