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Dadvar, A.

Publications and source records attributed to Dadvar, A..

2 recordsLinked to original sources

Diverse horizontally transferred cellulose biosynthesis gene clusters in Escherichia coli strains

The phosphoethanolamine modified exopolysaccharide cellulose is a major extracellular matrix component of Escherichia coli and Salmonella typhimurium. Upon enhanced acute virulence, however, cellulose production can be diminished or entirely abolished. Here, we report that homologs of the bcsABC core genes of the core genome bcs cellulose biosynthesis operon and even entire bcs operons can be mobilized on plasmids and are occasionally manifested on E. coli chromosomes at alternative locations. While BcsA2 and BcsA3 cellulose synthases are restricted to the genus Escherichia, the BcsA4 cellulose synthase and the entire associated bcs gene cluster are highly similar to one of the two chromosomal bcs gene clusters harboured by Klebsiella pneumoniae. Cyclic di-GMP turnover proteins known to post-translationally regulate cellulose biosynthesis are frequently localized on bcs operon bearing plasmids. In thermotolerant meat derived E. coli 740v1 harbouring a type 4 bcs operon on a plasmid, chemical and genetic evidence indicates that cellulose is produced in M9 minimal medium and upon activation by the second messenger cyclic di-GMP, an allosteric activator of the cellulose synthase, respectively. With gene duplication and horizontal gene transfer to contribute to the multiplication of bcs operons in a number of different bacterial species, the ecological role(s) of multiplication, transfer and replacement of cellulose biosynthesis operons also in species other than E. coli still need to be unraveled.

genomics↗

FAST-STEM: A human pluripotent stem cell engineering toolkit for rapid design-build-test-learn development of human cell-based therapeutic devices

Very recent clinical advances in stem cell derived tissue replacement and gene therapy, in addition to the rise of artificial intelligence-aided scientific discovery, have placed the possibility of sophisticated human cell-based therapies firmly within reach. However, development of such cells and testing of their engineered gene circuit components, has proven highly challenging, due to the need for generating stable cell lines for each design-build-test-learn engineering cycle. Current approaches to generating stable human induced pluripotent stem cell (hiPSC) lines are highly time-consuming and suffer from lack of control, poor integration efficiency, and limited functionality. Validation in clinically relevant stem cell derived tissues is also broadly lacking. Such drawbacks are prohibitive to repeatably conducting cutting-edge stem cell engineering with broad application within a realistic timeframe and will not scale with the future of regenerative medicine. We have developed FAST-STEM (Facile Accelerated Stem-cell Transgene integration with SynBio Tunable Engineering Modes), a hPSC engineering platform that drastically reduces the time to generate differentiation ready stem cell lines from several weeks to 5 days, exhibiting a ~612-fold improvement in transgene integration rate over previous methodologies. Additional FAST-STEM innovations include: (i) rapid and highly efficient transgene integration; (ii) copy number control; (iii) simultaneous or consecutive integration of multiple gene cassettes; (iv) library screen capability. In addition to this unique functional versatility, platform transportability and broad use case for stem cell-engineering was confirmed by differentiation into eight different cell types across nine different laboratories. This platform dramatically lowers the bar for integration of synthetic biology with regenerative medicine, enabling experiments which were previously deemed logistically impossible, thus paving the way for sophisticated human cell device development.

bioengineering↗