According to one embodiment, a vector set includes a first vector and a second vector. The first vector includes a transposase target sequence, a first promoter sequence ligated to downstream of the transposase target sequence, and a first reporter gene ligated to downstream of the first promoter sequence. The second vector includes a 5′-side transposase recognition sequence, a 3′-side transposase recognition sequence, and a first enhancer sequence arranged therebetween.

A chimeric antigen receptor (CAR) cell library is established through the fusion of a cell and a vector assembly. The vector assembly carries three genetic elements corresponding to a plurality of first genetic elements encoding one or more idiotype CARs, a second genetic element carrying one or more genetic circuits, and a third genetic element encoding one or more inducible proteins, respectively. The one or more genetic circuits are pre-programmed and are each a combination of a cis-regulatory factor and a transcription factor; and the one or more inducible proteins include one or two selected from the group consisting of a drug resistance protein and a suicide protein. By designing a CAR library-genetic circuit-inducible protein coupling scheme, the cell library construction and screening for complex and unknown disease target antigens are realized, such as to solve the problems that there are complex, diverse, and variable antigens.

Methods and systems for identifying and/or quantifying polypeptide binding interactions of ligand-binding polypeptides are disclosed. Detailed methods include methods for identifying binding ligands of ligand-binding polypeptides and methods for assessing changes in binding behavior due to alterations of ligand-binding polypeptides. Detailed systems include array-based systems that permit detection of ligand binding interactions at single-analyte resolution.

Methods and systems for identifying and/or quantifying polypeptide binding interactions of ligand-binding polypeptides are disclosed. Detailed methods include methods for identifying binding ligands of ligand-binding polypeptides and methods for assessing changes in binding behavior due to alterations of ligand-binding polypeptides. Detailed systems include array-based systems that permit detection of ligand binding interactions at single-analyte resolution.

The present disclosure provides methods for testing whether a target protein regulates viral RNA translation. The methods comprise a) introducing into a test host cell (where the test host cell comprises a catalytically inactive or a catalytically active CRISPR/Cas effector polypeptide): i) a reporter nucleic acid comprising a nucleotide sequence encoding a bicistronic translation monitor; and ii) a regulatory nucleic acid comprising a nucleotide sequence encoding a single guide RNA (sgRNA) that comprises a targeting sequence that specifically binds to a target sequence within a nucleic acid encoding the target protein; and b) detecting expression of the reporter proteins to determine whether a target protein regulates viral RNA translation via a Cap-dependent element or a Cap-independent element based on the expression of the reporter proteins in the test host cell as compared to the expression in the control host cell. Kits for conducting the methods disclosed herein are also provided.

The present invention provides a method for screening for a telomerase reverse transcriptase (TERT) phosphorylation inhibitor using TERT or a fragment thereof, and a TERT kinase.

RNA editing tools for use in systems designed to measure RNA in vivo and manipulate specific cell types are disclosed herein. An RNA sensor system comprising a) a single-stranded RNA (ssRNA) sensor comprising a stop codon and a payload; optionally wherein the ssRNA sensor further comprises a normalizing gene; and b) an adenosine deaminase acting on RNA (ADAR) deaminase; wherein the sensor is capable of binding to a ssRNA target to form a double-stranded RNA (dsRNA) duplex that becomes a substrate for the ADAR deaminase; wherein the substrate comprises a mispairing within the stop codon; and wherein the mispairing is editable by the ADAR deaminase, which editing can effectively remove the stop codon so as to enable translation and expression of the payload. A method of quantifying ribonucleic acid (RNA) levels using the RNA sensor system is also disclosed.

Systems and methods for identifying a current reprogramming status and for predicting a future reprogramming status for reprogramming intermediate cells (i.e., somatic cells undergoing reprogramming) are provided. Label-free autofluorescence measurements are combined with machine learning techniques to provide highly accurate identification of current reprogramming status and prediction of future reprogramming status. The identification of current reprogramming status utilizes metabolic endpoints from the autofluorescence data set. The prediction of future reprogramming status utilizes a pseudotime line constructed from autofluorescence data of reprogramming intermediate cells having a known reprogramming status.

The present invention provides a method of identifying a target for preparing an inhibitory chimeric antigen receptor (iCAR) or a protective chimeric antigen receptor (pCAR) capable of preventing or attenuating undesired activation of an effector immune cell. Also provided are a list of iCAR targets, as well as vectors and transduced effector immune cells comprising the nucleic acid molecule and methods for treatment of cancer comprising administering the transduced effector immune cells are further provided.

The present invention is related to a method for detecting and identifying protein-protein or protein-small molecule interactions using a bait and prey system. It is also related to bait and prey proteins, small molecules and constructs that are used for the methods described herein.