# US 12,191,002 B2 · Dynamic characterization of synthetic genetic circuits in living cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,191,002 B2 - Document ID: US12191002B2 - Status: Granted - Kind: B2 - Application number: 18/169,178 - Filed: 2023-02-14 - Published: 2023-09-07 - Granted: 2025-01-07 - Earliest claimed: 2018-06-28 - Assignees: Microsoft Technology Licensing, LLC - Applicants: Microsoft Technology Licensing, LLC - Inventors: Neil Dalchau; Andrew Nicholas John Brojer Phillips; Paul Grant - Web page: https://patentgrep.com/patent/US12191002B2 ## CPC - G16B 5/30: Dynamic-time models - G16B 40/10: Signal processing, e.g. from mass spectrometry [MS] or from PCR - C12N 15/63: Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression - G16B 5/00: ICT specially adapted for modelling or simulations in systems biology, e.g. gene-regulatory networks, protein interaction networks or metabolic networks - G16B 20/00: ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations - G16B 25/00: ICT specially adapted for hybridisation; ICT specially adapted for gene or protein expression - G16B: BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention relates to a method for determining one or more intrinsic properties of a DNA component from a plurality of measurements obtained over a time period from a cell culture, with each cell comprising the DNA component, wherein the DNA component is involved in transcription of one or more target signals, wherein the plurality of measurements comprises measurements relating to the density of the cell culture over the time period and measurements relating to the amount of the one or more target signals in the cell culture over the time period. ## Claims ### Claim 1 A method for determining an intrinsic property of a DNA component involved in transcription of a target signal, the method comprising: accessing a plurality of measurements obtained over a time period from a cell culture, wherein each cell in the cell culture comprises the DNA component, and wherein the plurality of measurements comprises a first measurement relating to an amount of the target signal in the cell culture over the time period and a second measurement relating to a density of the cell culture of the time period; estimating parameters quantifying the intrinsic property of the DNA component by: embedding the estimated parameters within a mathematical model that describes a production of the target signal over the time period; and minimizing a measure of a difference between outputs of the mathematical model and the first measurement; estimating parameter values for a second mathematical model that describes cell culture density over time by minimizing a measure of a difference between an output of the second mathematical model and the second measurement; and based on the estimated parameters and the estimated parameter values, identifying the intrinsic property. ### Claim 2 (depends on 1) The method according to claim 1, wherein each cell in the cell culture further comprises a reference promoter that initiates transcription of a reference signal. ### Claim 3 (depends on 2) The method according to claim 2, wherein the plurality of measurements comprises: measurements relating to amount of the reference signal in the cell culture over the time period. ### Claim 4 (depends on 3) The method according to claim 3, wherein the DNA component is an enhancer, and wherein the intrinsic property is an ability to recruit a polymerase enzyme. ### Claim 5 (depends on 3) The method according to claim 3, further comprising: estimating parameter values for a third mathematical model that describes a capacity of the cell culture to produce the reference signal over time, by minimizing a measure of a difference between an output of the third mathematical model and the measurements relating to the amount of the reference signal in the cell culture over the time period; and based on the estimated parameters for the second mathematical model, the estimated parameters of the third mathematical model, and the estimated parameter values, identifying the intrinsic property. ### Claim 6 (depends on 5) The method according to claim 5, wherein the third mathematical model models the capacity of the cell culture to produce the reference signal over the time period as a chemical reaction network. ### Claim 7 (depends on 1) The method according to claim 1, wherein minimizing the measure of the difference between outputs of the mathematical model and the first measurement uses a Markov chain Monte Carlo method. ### Claim 8 (depends on 1) The method according to claim 1, wherein the DNA component is a promoter, and wherein the intrinsic property is an ability to bind transcription factors. ### Claim 9 (depends on 1) The method according to claim 1, wherein the cell culture is a culture of bacterial, yeast or mammalian cells. ### Claim 10 (depends on 1) The method according to claim 1, wherein the target signal is a target protein. ### Claim 11 (depends on 10) The method according to claim 10, wherein the target protein is a fluorescent protein, and wherein the first measurement relating to the amount of the target signal in the cell culture over the time period are fluorescence measurements. ### Claim 12 (depends on 1) The method according to claim 1, further comprising: obtaining the plurality of measurements over the time period from the cell culture; and adapting the DNA component in vitro based on the one or more intrinsic properties. ### Claim 13 A system for determining an intrinsic property of a DNA component involved in transcription of a target signal, the system comprising: a processor; and a memory comprising computer-executable instructions that, when executed by the processor, cause the processor to perform the following operations: accessing a plurality of measurements obtained over a time period from a cell culture, wherein each cell in the cell culture comprises the DNA component, and wherein the plurality of measurements comprises a first measurement relating to an amount of the target signal in the cell culture over the time period and a second measurement relating to a density of the cell culture of the time period; estimating parameters quantifying the intrinsic property of the DNA component by: embedding the estimated parameters within a mathematical model that describes a production of the target signal over the time period; and minimizing a measure of a difference between outputs of the mathematical model and the first measurement; estimating parameter values for a second mathematical model that describes cell culture density over time by minimizing a measure of a difference between an output of the second mathematical model and the second measurement; and based on the estimated parameters and the estimated parameter values, identifying the intrinsic property. ### Claim 14 (depends on 13) The system according to claim 13, wherein each cell in the cell culture further comprises a reference promoter that initiates transcription of a reference signal. ### Claim 15 (depends on 14) The system according to claim 14, wherein the plurality of measurements comprises: measurements relating to an amount of the reference signal in the cell culture over the time period. ### Claim 16 (depends on 14) The system according to claim 14, wherein the memory further comprises computer-executable instructions that, when executed by the processor, cause the processor to perform one or more of the following operations: estimating parameter values for a third mathematical model that describes a capacity of the cell culture to produce the reference signal over time, by minimizing a measure of a difference between an output of the third mathematical model and the measurements relating to the amount of the reference signal in the cell culture over the time period. ### Claim 17 A computer-readable storage device comprising computer-executable instructions for determining an intrinsic property of a DNA component involved in transcription of a target signal, the computer-executable instructions when executed by a processor, cause the processor to perform the following operations: accessing a plurality of measurements obtained over a time period from a cell culture, wherein each cell in the cell culture comprises the DNA component, and wherein the plurality of measurements comprises a first measurement relating to an amount of the target signal in the cell culture over the time period and a second measurement relating to a density of the cell culture of the time period; estimating parameters quantifying the intrinsic property of the DNA component by: embedding the estimated parameters within a mathematical model that describes a production of the target signal over the time period; and minimizing a measure of a difference between outputs of the mathematical model and the first measurement; estimating parameter values for a second mathematical model that describes cell culture density over time by minimizing a measure of a difference between an output of the second mathematical model and the second measurement; and based on the estimated parameters and the estimated parameter values, identifying the intrinsic property. ### Claim 18 (depends on 17) The computer-readable storage device according to claim 17, wherein each cell in the cell culture further comprises a reference promoter that initiates transcription of a reference signal. ### Claim 19 (depends on 18) The computer-readable storage device according to claim 18, wherein the DNA component is an enhancer, and wherein the intrinsic property is an ability to recruit a polymerase enzyme. ### Claim 20 (depends on 17) The computer-readable storage device according to claim 17, wherein the computer-readable medium further comprises computer-executable instructions that, when executed by the processor, cause the processor to perform the following operations: estimating parameter values for a third mathematical model that describes cell culture density over time, by minimizing a measure of a difference between an output of the third mathematical model and measurements relating to a density of the cell culture over the time period.