33 results about "Gene Circuits" patented technology

Described are methods and compositions for the detection of target genes. The inventors have developed a synthetic nucleic acid sensor-effector gene circuit. In cells without a target gene, the circuit suppresses e.g., effector production, but in the presence of the target gene the suppression is subject to competition, such that the synthetic sensor is de-repressed and permits expression of the effector gene. The methods and compositions described further permit the selective expression of an effector gene in those cells expressing the target gene. In this manner, cells expressing a target gene can be selectively targeted for treatment or elimination. In certain aspects, the methods and compositions described permit the selective expression of an agent such as a therapeutic gene product, in a specifically targeted population of cells in an organism.

The invention discloses an iterative gene circuit based on a vibrio fischeri quorum sensing system and a T7 expression system. The iterative gene circuit comprises a signal molecular protein gene luxIof vibrio fischeri, a receptor protein gene luxR, a promoter sequence PluxI(1) for sensing a signal molecular and receptor protein complex, a T7 RNA polymerase gene T7 RNApoly and a green fluorescentprotein egfp for representing the circuit intensity. The gene circuit does not rely on an inducer and can utilize the T7 expression system to spontaneously and forcefully initiate target gene expression in any host bacterium; and gene elements are constructed on different expression vectors, the iterative gene circuit is constructed in engineering bacteria, the T7 expression system is utilized toamplify signals of the vibrio fischeri quorum sensing system, and the iterative gene circuit has the function of utilizing the T7 expression system to spontaneously and forcefully initiate target gene expression, and has a lower leakage ratio and higher initiating strength.

The invention provides a carrier assembly carrying a gene element combination, a recipient cell library, a preparation and screening method and an application, the recipient cell library is formed byfusing cells and the carrier assembly, and the carrier assembly at least carries three gene elements, respectively a plurality of first gene elements encoding one or more idiotypic synNotch receptors,respectively; a second gene element carrying one or more gene loops; a third gene element encoding one or more idiotypic chimeric antigen receptors. Wherein, when the first genetic element encodes one idiotypic synNotch receptor, the third genetic element must encode at least three idiotypic chimeric antigen receptors, and when the third genetic element encodes one idiotypic chimeric antigen receptor, the first genetic element must encode three idiotypic synNotch receptors. The gene loop is pre-programmed, a regulatory homeopathic factor is combined with a transcription factor, upon activation of the synNotch receptor encoded by the first gene element, the chimeric antigen receptor encoded by the third gene element is controllably expressed.

The present invention relates to a method of screening a target metabolite-overproducing mutant strain using a synthetic suicide genetic circuit, and more particularly to a method of screening only a metabolite-overproducing mutant strain while killing a mutant strain that does not produce the metabolite, by introducing into mutant strains a synthetic suicide genetic circuit comprising a suicide gene coupled with a riboswitch. The method for screening a target metabolite-overproducing mutant strain according to the present invention has advantages in that a metabolite-overproducing mutant strain having a relatively fast or slow growth rate can be separated by visual observation, and in that the riboswitch that is used in the synthetic suicide genetic circuit can be replaced depending on the kind of target metabolite, and thus the synthetic suicide genetic circuit can be applied commonly to various strains.

The invention relates to the technical field of biology, and discloses a single-input multi-output gene loop. The gene loop selects two pairs of inhibitory transcription factors having good orthogonality and remarkable inhibition efficiency differences and DNAs combined with the inhibitory transcription factors for responding to the cell surface antigen density, and different functional proteins are driven to be expressed according to the intensity of input signals. Effect cells are improved through the single-input multi-output gene loop, and can be driven to differentially express differentfunctional proteins such as cell factors along with density changes of antigens of target cells, and a combination effect on the target cells is achieved. According to the single-input multi-output gene loop, the efficiency for cell treatment of a single antigen can be improved, and the single-input multi-output gene loop is considered in a method for cell treatment of multiple antigens.

The invention provides a CRISPR-Cas9-based nucleic acid detection system and application thereof. A protein expression regulating and control system comprises Cas9 protein, RsgRNA and a cell-free protein expression system; and the RsgRNA comprises antisense nucleic acid and sgRNA according to a direction of 5' to 3'. The nucleic acid detection system provided by the invention combines a CRIPSR / Cas9 nucleic acid identification regulating and control system with a gene loop, regulates and controls the expression of protease by the RsgRNA, converts a nucleic acid signal into generation of the protease, and realizes rapid detection of the nucleic acid by detecting the protease.

The invention discloses a far-red light gene circuit expression control system. The far-red light gene circuit expression control system comprises a photoreceptor for sensing a far-red-light light source, a processing device for processing a signal transmitted by the photoreceptor and an effector for responding the signal transmitted by the processing device. The invention further discloses a gene circuit remote regulation and control system; the gene circuit remote regulation and control system comprises the far-red light gene circuit expression control system, a control device for sending out a remote control instruction and a far-red-light light source device. The invention further discloses application of the far-red light gene circuit expression control system and the gene circuit remote regulation and control system to treatment of diabetes mellitus. The invention further discloses a eukaryotic expression vector, an engineered cell or an engineered cell transplanting vector, which contains the far-red light gene circuit expression control system. The far-red light gene circuit expression control system can be used for rapidly regulating and controlling gene expression and has the characteristics of ultra-remote control, intelligent regulation and control of gene expression quantity, high regulating and controlling expression times, high time-space specificity, strong tissue penetrating power, good insulating property, no toxic or side effect and the like.

The invention discloses a far-red light gene circuit expression control system. The far-red light gene circuit expression control system comprises a photoreceptor for sensing a far-red-light light source, a processing device for processing a signal transmitted by the photoreceptor and an effector for responding the signal transmitted by the processing device. The invention further discloses a gene circuit remote regulation and control system; the gene circuit remote regulation and control system comprises the far-red light gene circuit expression control system, a control device for sending out a remote control instruction and a far-red-light light source device. The invention further discloses application of the far-red light gene circuit expression control system and the gene circuit remote regulation and control system to treatment of diabetes mellitus. The invention further discloses a eukaryotic expression vector, an engineered cell or an engineered cell transplanting vector, which contains the far-red light gene circuit expression control system. The far-red light gene circuit expression control system can be used for rapidly regulating and controlling gene expression and has the characteristics of ultra-remote control, intelligent regulation and control of gene expression quantity, high regulating and controlling expression times, high time-space specificity, strong tissue penetrating power, good insulating property, no toxic or side effect and the like.

The invention discloses a gene circuit simulating an artificial neural network. The gene circuit comprises at least one input layer, at least two hidden layers and at least one output layer; the inputlayer comprises a plurality of input layer nodes; each hidden layer comprises a plurality of hidden layer nodes, the output layer comprises a plurality of output layer nodes, an input vector of eachinput layer node is a promoter and an input gene, each hidden layer node comprises a hidden layer gene, and each hidden layer gene is regulated and controlled by a promoter of the gene of the layer ora gene product of the gene of the previous layer; the output node outputs a gene circuit result; the regulation and control of the product of the previous layer of gene on the hidden layer gene and the output layer node accord with an excitation function; and the input layer node is regulated and controlled by the input promoter to accord with an excitation function. By implementing the method, the artificial neural network can be simulated by adopting a gene circuit structure, so that complex operation is simulated on a molecular level, and complex application in the fields of biology, medicine, chemistry, electronics and the like is realized.

A Gene circuit based on lung-cancer marker detection consists of a marker single-index detection module and a fusion circuit module in sequential connection; the marker single-index detection module consists of an upstream branch and a downstream branch; in the upstream branch, a marker starting circuit Input-XY and an inverter Z-Output are in sequential connection; in the downstream branch, a marker starting circuit Input-Y, an inverter Y-W, an inverter W-Z and an inverter Z-Output are in sequential connection; and the fusion circuit module is formed by sequentially connecting an inverter ABC-G with three-port input and an inverter G-GFP. The gene circuit combines the advantages of the two modules, and is capable of realizing rapid response and logic determination, and the detection and alarm effects of the lung cancer marker are realized. The gene circuit has relatively good practical value and wide application prospect in the technical field of synthetic biology.

The invention provides a cancer cell state identification gene circuit group and a preparation method thereof, and belongs to the technical field of gene engineering. The cancer cell state identification gene circuit group comprises a first identification gene circuit and a second identification gene circuit which are both used for identifying cancer cells in the same cancer cell state. A microRNA sequence is inserted into the 3 'UTR segment of the positive promoter, and the positive promoter, the negative promoter and the microRNA are from cancer cells in a cancer cell state. The invention also discloses a preparation method of the cancer cell state identification gene circuit group. The positive promoter, the negative promoter and the microRNA capable of representing different phenotypes of tumor cells are obtained, so that the first identification gene circuit and the second identification gene circuit are formed, and the bar code area capable of assisting judgment is combined, so that the cancer cell state can be accurately calibrated, and the accuracy of cancer cell detection is improved. And the proportion information of the cancer cells in various states can be analyzed by means of high-throughput cell experiments and the like, so that the analysis of heterogeneity among the cancer cells is facilitated.

The invention relates to a gene circuit structure for realizing glioma marker detection. The gene circuit structure consists of a single-index detection module and an integration circuit module. The two modules are in sequential connection mutually. The single-index detection module is composed of a phase inverter, which consists of an input end, an output end and a reverse module and is used for negating an input signal and outputting a reverse signal. The integration circuit is composed of a three-port input NAND gate, which is used for performing NAND operation on the input. When the three inputs are all at a high level, a low level signal is output by the circuit. The gene circuit structure provided by the invention has the advantages of simple structure, fast response, stable output and good performance, overcomes the interference from independent work of a single module and other unfavorable factors, and realizes glioma marker detection and alarming functions. The gene circuit structure has good practical value and broad application prospects in the technical field of synthetic biology.

Provided are: a selection method for a gene switch and a gene circuit, including using, as a selector, an expression vector containing at least a gene sequence whose expression is controlled by a transcription regulatory factor to be expressed when a genetic switch and a genetic circuit including the genetic switch operate, and a promoter sequence operably linked to the gene sequence upstream thereof; and an expression vector to be used in the selection method. This enables an effective selection method for a genetic switch and a genetic circuit, the selection method being able to be conducted within a short time period and with high selection efficiency and less leakiness.

Aspects of the present disclosure provide a platform that triggers potent and effective immunotherapy against tumors from within tumors themselves, thus overcoming limitations of existing cancer immunotherapies and tumor-detecting gene circuits.

The invention provides a chimeric antigen receptor cell library carrying a gene element combination, preparation and screening methods and application. The chimeric antigen receptor cell library is formed by fusion of cells and a carrier assembly, the carrier assembly carries three gene elements, and the three gene elements are respectively as follows: a plurality of first gene elements for encoding one or more unique chimeric antigen receptors; a second genetic element carrying one or more genetic loops; and a third genetic element encoding one or more inducible proteins. The genetic loop ispreprogrammed and is a combination of a regulatory homeopathic factor and a transcription factor; and the inducible protein comprises one or two of drug resistance protein and suicide protein. By designing the chimeric antigen receptor library-genetic loop-inducible protein coupling scheme, the cell library and screening of complex and unknown disease target antigens are realized, so that the problems that antigens in abnormal antigen expression diseases are complex, diverse and easy to change, target identification is difficult and the like are solved. The method has a wide application prospect.

The invention discloses a recombinant plasmid, a recombinant engineering bacterium as well as a construction method and application thereof in improving the yield of D-psicose, and belongs to the technical field of enzyme directed evolution. The invention designs a directed evolution recombinant plasmid, and the directed evolution recombinant plasmid can be regarded as a gene circuit, namely, a gene circuit with DTE enzyme efficiency. At the same time, the invention also discloses the recombinant engineering bacterium constructed based on the recombinant plasmid, specifically E. coli DH5alphapSB1C3-psiR-pPsi-EGFP-Cp-KanR-DTE. The gene circuit of the present invention uses a repressor protein PsiR and a corresponding promoter pPsi to realize a biosensor of D-psicose; and the bacterium can sense the production of D-psicose in the body to express different levels of antibiotics resistance genes, so that the efficiency of DTE enzyme is transformed into survival advantages of bacterium.

The invention provides an AND gate gene circuit and a method for obtaining the AND gate gene circuit, and belongs to the technical field of gene engineering. The AND gate gene circuit comprises a first end sequence, a guide sequence and a second end sequence which are connected in sequence; the first terminal sequence comprises a CasRx-C terminal sequence and a first promoter which are connected in sequence; the guide sequence at least comprises a guide promoter and a gRNA sequence which are connected in sequence; andthe second terminal sequence at least comprises a second promoter and a CasRx-N terminal sequence, and resolution sites of the CasRx-C terminal sequence and the CasRx-N terminal sequence are located in a secondary structure hinge region of the three-dimensional prediction structure of the amino acid sequence of CasRx. The invention further discloses a method for obtaining the AND gate gene circuit. When the first promoter and the second promoter are different promoters, the CasRx fused again can exert a gene editing effect only under the precondition that the two promoters can normally exert functions by utilizing the characteristics similar to those of an AND gate circuit, so that whether the two promoters interfere with each other or not can be quickly judged.

The invention provides a code-level cerebral apoplexy diagnosis gene circuit design method. The method comprises the following steps: 1, building a gene circuit element model base, wherein the elementmodel library comprises circuit models for realizing three basic logic functions of AND, OR and NOT; 2, performing logic abstraction on a marker or marker group detection function by using program design, and outputting a detection circuit function logic expression; 3, converting the logic expression into a gate-level circuit diagram, and laying a foundation for building of a gene circuit in thenext step; 4, detecting a single marker or a plurality of markers of cerebral apoplexy to realize disease diagnosis; through the above steps, design and construction of different logic function detection circuits of the cerebral stroke marker and the marker group are realized, and real-time detection and judgment are performed on biological marker molecules, so the circuit achieves early warning and diagnosis effects on cerebral stroke, and meanwhile, the circuit design efficiency and the detection accuracy are improved.

The invention discloses a far-red light gene circuit expression control system. The far-red light gene circuit expression control system comprises a photoreceptor for sensing a far-red-light light source, a processing device for processing a signal transmitted by the photoreceptor and an effector for responding the signal transmitted by the processing device. The invention further discloses a gene circuit remote regulation and control system; the gene circuit remote regulation and control system comprises the far-red light gene circuit expression control system, a control device for sending out a remote control instruction and a far-red-light light source device. The invention further discloses application of the far-red light gene circuit expression control system and the gene circuit remote regulation and control system to treatment of diabetes mellitus. The invention further discloses a eukaryotic expression vector, an engineered cell or an engineered cell transplanting vector, which contains the far-red light gene circuit expression control system. The far-red light gene circuit expression control system can be used for rapidly regulating and controlling gene expression and has the characteristics of ultra-remote control, intelligent regulation and control of gene expression quantity, high regulating and controlling expression times, high time-space specificity, strong tissue penetrating power, good insulating property, no toxic or side effect and the like.

The invention discloses a blue-light-controlled dynamic regulation system and its application, specifically discloses a blue-light-controlled dynamic regulation system and its application in the production of acetoin, belonging to the technical field of bioengineering. The present invention selects the blue light photosensitive protein by means of molecular biology, utilizes the light-sensitive protein’s specific response to the light source characteristic of a specific wavelength and the advantage of not needing to add an inducer from an external source, and introduces the dynamic regulation of targeting FtsZ+FtsA into engineering Escherichia coli The gene circuit realizes the production of acetoin in the inorganic salt medium. The yield of acetoin produced by the invention can reach 63g / L, and the conversion rate can reach 0.57g / g glucose.

The invention provides a cancer cell analysis gene circuit assembly and a production method thereof, and belongs to the technical field of gene engineering. The cancer cell analysis gene circuit assembly comprises a first identification gene circuit and a second identification gene circuit which are both used for identifying cancer cells in the same cancer cell state. A microRNA sequence is inserted into the 3'UTR segment, and the microRNA is from cancer cells in the cancer cell state. The invention also discloses the production method of the cancer cell analysis gene circuit assembly. A positive synthetic promoter and a negative synthetic promoter are formed by obtaining a positive enhancer and a negative enhancer capable of representing different phenotypes of the tumor cells, and the bar code area capable of assisting judgment is combined, so that the cancer cell state can be accurately calibrated, the proportion information of the cancer cells in various states can be analyzed by means of high-throughput cell experiments and the like, and the analysis of heterogeneity among the cancer cells is facilitated.

A proximity dependent split T7 RNAP (RNA polymerase) sensor using continuous molecular evolution is described. The versatility of the platform is described by creating robust light and small molecule-responsive genetic sensors. The activity-responsive RNAP platform dramatically simplifies and expands genetic circuit creation, and opens new opportunities in protein engineering, synthetic biology, and bioengineering.

The invention relates to a temperature-sensitive gene loop system as well as a construction method and application thereof. According to the invention, a series of temperature-sensitive biosensors suitable for B. subtilis are designed and reconstructed; a bidirectional gene loop is constructed based on a CRISPRi system, the bidirectional gene loop is combined with a single-input multi-output gene loop to construct a high-version single-input multi-output gene loop, and activation expression or / and down-regulation expression of a plurality of metabolism modules at different fermentation time points is / are realized. And then dynamically regulating and controlling three modules participating in metabolic synthesis of 2 '-FL by using the obtained gene loop: a 2'-FL synthesis module, a cofactor supply module and a competitive pathway module. The cell growth stage and the 2 '-FL synthesis stage can be decoupled by dynamic regulation and control through a gene loop, the metabolic pressure of host cells is relieved, metabolic flux is reasonably distributed, and efficient synthesis of 2'-FL is achieved.

The invention discloses a far-red light gene circuit expression control system. The far-red light gene circuit expression control system comprises a photoreceptor for sensing a far-red-light light source, a processing device for processing a signal transmitted by the photoreceptor and an effector for responding the signal transmitted by the processing device. The invention further discloses a gene circuit remote regulation and control system; the gene circuit remote regulation and control system comprises the far-red light gene circuit expression control system, a control device for sending out a remote control instruction and a far-red-light light source device. The invention further discloses application of the far-red light gene circuit expression control system and the gene circuit remote regulation and control system to treatment of diabetes mellitus. The invention further discloses a eukaryotic expression vector, an engineered cell or an engineered cell transplanting vector, which contains the far-red light gene circuit expression control system. The far-red light gene circuit expression control system can be used for rapidly regulating and controlling gene expression and has the characteristics of ultra-remote control, intelligent regulation and control of gene expression quantity, high regulating and controlling expression times, high time-space specificity, strong tissue penetrating power, good insulating property, no toxic or side effect and the like.

The invention provides a method for obtaining a driving element, and belongs to the technical field of genetic engineering. The method for obtaining the driving element comprises the following steps: step 1, preparing a drug-resistant cancer cell subgroup under the condition of combined culture of a chemotherapeutic drug and a targeted drug; 2, preparing at least one preselected promoter; 3, constructing a promoter test lentivirus; 4, constructing a cell phenotype state evaluator; 5, obtaining a stably transferred cancer cell line; step 6, obtaining a dual stable cancer cell line; and step 7, screening based on the survival rate of the dual stable transformation cancer cell line to obtain the driving element. On the basis of the drug-resistant culture result of a double stably transferred cancer cell line, the type of the preselected promoter capable of highly recognizing various subtype cancer cell lines is judged, so that the preselected promoter can be used as the driving element in a subsequently constructed gene circuit.

The invention discloses a dynamic control system controlled by near-infrared light and its application, specifically discloses a dynamic control system related to combined control of near-infrared light and its application in the production of acetoin, which belongs to the technical field of metabolic engineering. The present invention selects the near-infrared photosensitive protein by means of molecular biology. Taking advantage of the photosensitive protein's specific response to light source characteristics of specific wavelengths and the advantages of not needing to add exogenous inducers, the production of acetoin in inorganic salt medium was realized by introducing a dynamic regulatory gene circuit targeting RpoS into engineered Escherichia coli. The yield of acetoin produced by the invention can reach 66.8g / L, and the conversion rate can reach 0.58g / g glucose.

The invention discloses an establishing method of a logic and gate gene circuit for detecting bladder cancer. The method includes the steps of firstly, amplifying an hUPPII promoter sequence shown in SEQ ID NO:1, an aminoacyl tRNA synthetase AcKRS gene sequence shown in SEQ ID NO:2, an hTERT promoter sequence shown in SEQ ID NO:3, a tRNA sequence shown in SEQ ID NO:4 and a pCMV6-AC-GFP37TAG plasmid sequence shown in SEQ ID NO:6 through a PCR method; secondly, connecting an hUPPII promoter, an AcKRS gene, an hTERT promoter and tRNA into a psiCHECK-2 plasmid shown in SEQ ID NO:7 to establish a recombinant vector; thirdly, transplanting the recombinant carrier and a pCMV6-AC-GFP37TAG plasmid into bladder cancer cells to establish the gene circuit. When the gene circuit is transplanted into bladder cancer cells and acetylated lysine is added, green fluorescent proteins are only expressed in bladder cancer cells instead of being expressed in normal cells.