The present disclosure provides methods for genetically modifying lymphocytes and methods for performing adoptive cellular therapy that include transducing T cells and/or NK cells. The methods can include inhibitory RNA molecule(s) and/or engineered signaling polypeptides that can include a lymphoproliferative element, and/or a chimeric antigen receptor (CAR), for example a microenvironment restricted biologic CAR (MRB-CAR). Additional elements of such engineered signaling polypeptides are provided herein, such as those that drive proliferation and regulatory elements therefor, as well as replication incompetent recombinant retroviral particles and packaging cell lines and methods of making the same. Numerous elements and methods for regulating transduced and/or genetically modified T cells and/or NK cells are provided, such as, for example, those including riboswitches, MRB-CARs, recognition domains, and/or pH-modulating agents.

A method screens for aptamers by using a microarray microfluidic chip. The screening chip integrates microarray and microfluidic technology to integrate the positive and negative screening process on a microfluidic chip, and obtains aptamers with high affinity after 7 rounds of screening. It also discloses specific steps for screening of lactoferrin aptamers, including detailed processes such as chip preparation, positive and negative screening processes, and PCR amplification. The aptamers screened by the method have good specificity and affinity to the target protein. The aptamers are easier to be obtained than the antibody, and can be synthesized rapidly in large quantities in vitro. The preparation method is simpler and faster, so aptamers are expected to be a useful complement to antibody technology in many areas.

Methods of selecting an aptamer that specifically binds to a target molecule complexed with a derivatization agent. Also disclosed are specific aptamers and methods of use thereof.

Polynucleotides, such as aptamers, comprising at least first one 5-position modified pyrimidine and at least one second 5-position modified pyrimidine are provided, wherein the first and second 5-position modified pyrimidines are different. Methods of selecting and using such polynucleotides, such as aptamers, are also provided.

The present disclosure describes compositions and methods for rapid selection of both binding and functional oligonucleotides (DNA, RNA, or any natural or synthetic analog of these). In certain embodiments, provided herein are flow cells (e.g., flow cells for an Illumina sequencing instrument or a Polonator sequencing instrument) comprising within its flow chamber a plurality of immobilized aptamer clusters (e.g., from an aptamer library described herein) and, optionally, one or more target cells (e.g., cancer cells, immune cells, etc.) and/or a detectable indicator of cellular function (e.g., a fluorescent indicator of apoptosis, cell proliferation, gene or protein expression, etc.). In certain embodiments, provided herein are methods of using such an aptamer cluster-containing flow cell to identify functional aptamers from an aptamer library (e.g., in a sequencing instrument, such as an Illumina sequencing instrument).

The present disclosure provides methods, kits and compositions for identifying nucleic acid agents having a desired property, e.g., a property of specifically binding to a target (such as a protein target) with high affinity. More specifically, the present disclosure provides methods, kits and compositions for identifying candidate nucleic acid agents with both high specificity and affinity for a target.

The present application discloses nucleic acid aptamer based signaling polynucleotides (SPNs) that specifically bind an allergen protein. Provided in the present invention include aptamers, SPNs, SPN-complement complexes, magnetic particle conjugates, DNA printed glass slides and detection agents, and detection methods using the same for detecting the presence, or absence of an allergen protein in a food sample.

Described herein is a reversible aptamer selection technology for production-scale isolation of label-free cells (e.g., CD8+ T cells). Provided herein are methods for isolating a cell of interest from a biological sample by contacting the biological sample with an aptamer that specifically binds a cell surface marker specific for the cell of interest; separating the aptamer-bound cells from cells not bound to the aptamer; and recovering a cell of interest by disrupting binding of the aptamer to the cell surface marker.

A method of producing an aptamer selectively binding a non-canonical structure of a target nucleic acid molecule includes the steps of: incubating a plurality of nucleic acid sequences with an enantiomer of the non-canonical structure under suitable conditions to obtain one or more candidate nucleic acid sequences binding to the enantiomer of the non-canonical structure, purifying and amplifying the one or more candidate nucleic acid sequences; repeating said incubating, purifying and amplifying steps for a predetermined number of cycles under different conditions; and producing an enantiomer for selected amplified candidate nucleic acid sequence to obtain the aptamer capable of selectively binding the non-canonical structure of the target nucleic acid molecule. An aptamer selectively binding to a non-canonical structure of a nucleic acid molecule or its enantiomer, the aptamer comprising a sequence of SEQ ID NO: 11; as well as uses of the aptamer or its enantiomer.

The present disclosure relates to a closed loop aptamer development system that identifies one or more aptamers observed experimentally and implements machine-learning models to identify other aptamers not observed experimentally. Particularly, aspects of the present disclosure are directed to receiving a query concerning one or more targets, acquiring a library of aptamers that potential satisfy the query, identifying a first set of aptamers from the library of aptamers that substantially or completely satisfy the query, obtaining sequence data for the first set of aptamers, generating, by a prediction model, a third set of aptamers derived from the sequence data for the first set of aptamers, validating the third set of aptamers that substantially or completely satisfy the query, and upon validating the third set of aptamers and in response to the query, providing the third set of aptamers as a result to the query.