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논문 기본 정보
- 자료유형
- 학술대회자료
- 저자정보
- 발행연도
- 2021.4
- 수록면
- 42 - 42 (1page)
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Living organisms have a variety of mechanisms to sense and respond to the environmental signals by dynamically regulating their genetic expression networks. Harnessing this ability, genetically encoded biosensors, mimicking natural regulation networks by assembling basic biological parts like promoter, ribosome binding site, operator, reporter etc. into genetic circuit, are developed to recognize the analytes and transduce their signals to a measurable output. Dose-response curve, mapping the genetic circuits to their function, shapes how individual biosensors respond to the specific signals, which is crucial not only for the specific usage scenario of genetic encoded biosensor, but also for illustrating their regulatory functions in living cells. In this report, we would like show our recent attempts on the application of a dose-response profiling method, namely fluorescence-activated cell sorting coupled with next-generation sequencing (FACS-seq), in generating accurate dose-response curves for thousands of biosensors in a massively parallel manner, which provides a powerful platform for dissecting the mechanistic basis of the regulatory elements in living cells, and for the fine tuning of biosensors in a customized and low-cost manner.
As the first example, we focused on tnaC, which encodes the tryptophan operon leader peptide in bacteria and is a model of macromolecular-machinery-dynamics-dependent regulatory elements. Working as a molecular sensor, tnaC responds to intracellular tryptophan (Trp) and regulates the biosynthesis of indol. We used FACS-seq to generate accurate response curves for all possible codon substitutions in tnaC. The FACS-seq results allowed us to generate comprehensive genotype-phenotype maps for 1,450 tnaC vari ... 전체 초록 보기
As the first example, we focused on tnaC, which encodes the tryptophan operon leader peptide in bacteria and is a model of macromolecular-machinery-dynamics-dependent regulatory elements. Working as a molecular sensor, tnaC responds to intracellular tryptophan (Trp) and regulates the biosynthesis of indol. We used FACS-seq to generate accurate response curves for all possible codon substitutions in tnaC. The FACS-seq results allowed us to generate comprehensive genotype-phenotype maps for 1,450 tnaC vari ... 전체 초록 보기
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