Cell-Based Biosensors Based on Intein-Mediated Protein Engineering for Detection of Biologically Active Signaling Molecules

Analytical Chemistry
Hyunjin JeonYoungeun Kwon

Abstract

Live-cell-based biosensors have emerged as a useful tool for biotechnology and chemical biology. Genetically encoded sensor cells often use bimolecular fluorescence complementation or fluorescence resonance energy transfer to build a reporter unit that suffers from nonspecific signal activation at high concentrations. Here, we designed genetically encoded sensor cells that can report the presence of biologically active molecules via fluorescence-translocation based on split intein-mediated conditional protein trans-splicing (PTS) and conditional protein trans-cleavage (PTC) reactions. In this work, the target molecules or the external stimuli activated intein-mediated reactions, which resulted in activation of the fluorophore-conjugated signal peptide. This approach fully valued the bond-making and bond-breaking features of intein-mediated reactions in sensor construction and thus eliminated the interference of false-positive signals resulting from the mere binding of fragmented reporters. We could also avoid the necessity of designing split reporters to refold into active structures upon reconstitution. These live-cell-based sensors were able to detect biologically active signaling molecules, such as Ca2+ and cortisol, as well...Continue Reading

References

Dec 1, 1995·Cell·E M BrownS C Hebert
Jul 8, 2000·Trends in Cell Biology·D Chin, A R Means
Feb 24, 2001·Biosensors & Bioelectronics·S S IqbalJ P Chambers
Jun 28, 2001·Physiological Reviews·A Aranda, A Pascual
Sep 11, 2002·Angewandte Chemie·Sergey V Dorozhkin, Matthias Epple
Aug 28, 2003·Journal of the American Chemical Society·Henning D MootzTom W Muir
Sep 30, 2004·Angewandte Chemie·Henning D MootzTom W Muir
Aug 29, 2007·Analytica Chimica Acta·Takeaki Ozawa
Jun 19, 2008·Analytica Chimica Acta·Xudong FanYuze Sun
Dec 17, 2008·Structure·Qi WangHolger Sondermann
Dec 23, 2008·The Journal of Biological Chemistry·Jasper AkerboomEric R Schreiter
Aug 21, 2009·Chembiochem : a European Journal of Chemical Biology·Hangxiang WangItaru Hamachi
Aug 27, 2009·Chembiochem : a European Journal of Chemical Biology·Henning D Mootz
Sep 23, 2009·Chemical Society Reviews·Wolf B FrommerRobert E Campbell
Feb 15, 2011·Molecular BioSystems·Muhammad Awais, Takeaki Ozawa
Jul 7, 2011·Proceedings of the National Academy of Sciences of the United States of America·Anca Dragulescu-AndrasiSanjiv S Gambhir
Sep 10, 2011·Science·Yongxin ZhaoRobert E Campbell
Mar 13, 2012·Journal of the American Chemical Society·Radhika BorraJulio A Camarero
Jun 28, 2012·Journal of the American Chemical Society·Neel H ShahTom W Muir
Jan 16, 2013·Molecular BioSystems·Deokho JungYoungeun Kwon
Feb 20, 2013·Chemical Society Reviews·Anthony P F Turner
Mar 20, 2013·Journal of the American Chemical Society·Neel H ShahTom W Muir
Mar 19, 2014·Chemical Science·Neel H Shah, Tom W Muir
Jun 7, 2014·Chemical Reviews·Qingjun LiuPing Wang
May 16, 2015·Chemical Communications : Chem Comm·Deokho JungYoungeun Kwon
Sep 1, 2015·Biosensors & Bioelectronics·Xian-En ZhangDianbing Wang
Sep 30, 2016·Scientific Reports·Natalia V BarykinaGrigori N Enikolopov

❮ Previous
Next ❯

Citations

Mar 11, 2020·Annual Review of Chemical and Biomolecular Engineering·Emily A BerckmanWilfred Chen
Mar 18, 2021·Biochimie·Sunita PandaSasmita Nayak
May 6, 2021·International Journal of Molecular Sciences·Jeahee RyuYoungeun Kwon
Apr 24, 2021·Journal of Biomedical Research·Mihaela Gheorghiu
Aug 17, 2019·Journal of the American Chemical Society·Josef A GramespacherTom W Muir
Sep 15, 2018·Bioconjugate Chemistry·Marissa A Morales, Jeffrey Mark Halpern

❮ Previous
Next ❯

Related Concepts

Related Feeds

CREs: Gene & Cell Therapy

Gene and cell therapy advances have shown promising outcomes for several diseases. The role of cis-regulatory elements (CREs) is crucial in the design of gene therapy vectors. Here is the latest research on CREs in gene and cell therapy.

Bioinformatics in Biomedicine

Bioinformatics in biomedicine incorporates computer science, biology, chemistry, medicine, mathematics and statistics. Discover the latest research on bioinformatics in biomedicine here.

Allogenic & Autologous Therapies

Allogenic therapies are generated in large batches from unrelated donor tissues such as bone marrow. In contrast, autologous therapies are manufactures as a single lot from the patient being treated. Here is the latest research on allogenic and autologous therapies.

Calcium & Bioenergetics

Bioenergetic processes, including cellular respiration and photosynthesis, concern the transformation of energy by cells. Here is the latest research on the role of calcium in bioenergetics.

Biosensor Development & Applications

Biosensor development and applications describe recent progress in biosensor development and their applications in the field of antibiotics and metabolic engineering. Discover the latest research on biosensor development and applications in this feed.