Brian Belmont

Provided herein are methods and compositions for single cell characterization using affinity-oligonucleotide conjugates. In some aspects, such methods may comprise attaching a first vessel barcoded polynucleotide to an oligonucleotide portion of an affinity-oligonucleotide conjugate, which binds to a target antigen expressed by a single cell that is isolated in a single vessel. In some aspects, the oligonucleotide portion of the affinity-oligonucleotide conjugate may comprise an antigen… Show more

Provided herein are methods and compositions for single cell characterization using affinity-oligonucleotide conjugates. In some aspects, such methods may comprise attaching a first vessel barcoded polynucleotide to an oligonucleotide portion of an affinity-oligonucleotide conjugate, which binds to a target antigen expressed by a single cell that is isolated in a single vessel. In some aspects, the oligonucleotide portion of the affinity-oligonucleotide conjugate may comprise an antigen identification sequence (AID). In some aspects, the oligonucleotide portion of the affinity-oligonucleotide conjugate may further comprise an affinity molecular barcode (AMB) sequence. In some aspects, such methods may further comprise lysing the single cell and attaching a second vessel barcoded polynucleotide to a cell polynucleotide from the single cell. Show less

Provided herein are methods and compositions for single cell characterization using affinity-oligonucleotide conjugates. Provided herein are methods and compositions for single cell charaterization using tetramer-oligonucleotide conjugates.

Provided herein are methods and composition for high-throughput T-cell receptor target identification of natively paired T-cell receptor sequences.

Provided herein are methods and compositions for single cell characterization using affinity-oligonucleotide conjugates, including tetramer-oligonucleotide conjugates (e.g., pMHC-oligonucleotide). In some aspects, such methods may comprise the steps of: forming a vessel comprising an affinity-oligonucleotide conjugate and a T cell; attaching a vessel barcoded polynucleotide or complement thereof to: (a) an oligonucleotide portion of a pMHC-oligonucleotide conjugate, and (b) a T cell receptor… Show more

Provided herein are methods and compositions for single cell characterization using affinity-oligonucleotide conjugates, including tetramer-oligonucleotide conjugates (e.g., pMHC-oligonucleotide). In some aspects, such methods may comprise the steps of: forming a vessel comprising an affinity-oligonucleotide conjugate and a T cell; attaching a vessel barcoded polynucleotide or complement thereof to: (a) an oligonucleotide portion of a pMHC-oligonucleotide conjugate, and (b) a T cell receptor (TCR) polynucleotide originating from the T cell; and sequencing the oligonucleotide portion or a complement thereof and the TCR polynucleotide or a complement thereof. Additional methods provided herein include determining the binding affinity of a T cell receptor (TCR) to a pMHC-oligonucleotide. Compositions of the present invention encompass a plurality of vessels, wherein a vessel of the plurality of vessels may comprise a single T cell; and a vessel barcoded polynucleotide, wherein the vessel may further comprise a pMHC-oligonucleotide conjugate. Show less

Provided are binding molecules, such as TCRs or antigen binding fragments thereof and antibodies and antigen-binding fragments thereof, such as those that recognize or bind human papilloma virus (HPV) 16, including HPV 16 E6 and HPV 16 E7. Also provided are engineered cells containing such binding molecules, compositions containing the binding molecules or engineered cells, and methods of treatment, such as administration of the binding molecules, engineered cells, or compositions.

Provided herein are methods and compositions for activated amplification of a single template polynucleotide molecule, such as for cell barcoding and DNA sequencing.

In vitro SELEX has been used to discover high affinity RNA aptamers interacting with the tetracycline repressor protein in a tetracycline-dependent manner. Using in silico RNA folding predictions to guide the design of both aptamer truncations and mutants, minimized tetracycline repressor protein high affinity binding aptamers have been defined. Using one such aptamer, inducible post-transcriptional regulation in vivo has been demonstrated that is predicated on a direct interaction between a… Show more

In vitro SELEX has been used to discover high affinity RNA aptamers interacting with the tetracycline repressor protein in a tetracycline-dependent manner. Using in silico RNA folding predictions to guide the design of both aptamer truncations and mutants, minimized tetracycline repressor protein high affinity binding aptamers have been defined. Using one such aptamer, inducible post-transcriptional regulation in vivo has been demonstrated that is predicated on a direct interaction between a tetracycline repressor protein and a RNA aptamer element. These aptamer components can be integrated in any organism to inducibly regulate RNA translation of a gene of interest. Show less

Provided are receptor tyrosine kinase-like orphan receptor 1 (ROR1)-binding molecules, in particular, to human antibodies specific for ROR1, including antibody fragments. The present disclosure further relates to recombinant receptors, including chimeric antigen receptors (CARs) that contain such antibodies or fragments, and polynucleotides that encode the antibodies, antigen-binding fragments or receptors specific for ROR1. The disclosure further relates to genetically engineered cells… Show more

Provided are receptor tyrosine kinase-like orphan receptor 1 (ROR1)-binding molecules, in particular, to human antibodies specific for ROR1, including antibody fragments. The present disclosure further relates to recombinant receptors, including chimeric antigen receptors (CARs) that contain such antibodies or fragments, and polynucleotides that encode the antibodies, antigen-binding fragments or receptors specific for ROR1. The disclosure further relates to genetically engineered cells, containing such ROR1 -binding proteins and receptors, and related methods and uses thereof in adoptive cell therapy. Show less