Methods for increasing expression of a target gene, the method comprising introducing a CCCTC-binding factor (CTCF) binding site (CTCF-BS) into a promoter region of the target gene, e.g., within 500, 250, 200, 150, 100, 50, or 25 nucleotides of the transcription start site (TSS) for the target gene, and optionally expressing in or introducing into the cell a CTCF protein or variant thereof.

Compositions and methods are provided for delivering hydrophilic functional agents to plants using lipid nanoparticles (NLPs). The NLPs comprise a hydrophilic core surrounded by a lipid shell including at least one sterol, one phospholipid, and one cationic or ionizable lipid. The hydrophilic core enables encapsulation and stabilization of water-soluble agents such as nucleic acids, peptides, small molecules, or plant hormones. The NLPs enhance uptake, protect cargo from degradation, and facilitate delivery to internal plant tissues, including meristems and vascular structures. Methods of treating plants and plant parts with the compositions are also provided.

Provided herein are methods, systems, and compositions for detecting and/or quantifying analytes. The methods, systems and compositions may comprise a first component of a signal-generating complex and a second component of signal-generating complex. The first component may be immobilized to a solid substrate via a disruptable linker at a first location of a reaction vessel and a second component may be immobilized to the solid substrate at a second location of the reaction vessel. The linker may be disrupted allowing the signal-generating complex to form.

The present invention is directed to a method for ex-vivo-engineering of cells, in particular stem cells or T cells, preferably hematopoictic stem and/or progenitor cells, mesenchymal stem cells, or T cells comprising a step of culturing the cells on a three-dimensional scaffold. The method of the invention is capable of improving the efficiency of genetic modification of cells and the functionality of the engineered cells.

Provided herein are systems of modulating gene expression, methods of use thereof, and cells engineered thereof for the purpose of differentiating cells, for example hematopoietic cells.

Disclosed herein are methods and compositions for increasing serine levels, increasing ceramide levels, and/or decreasing deoxyceramide, such as deoxydihydroceramide, levels in one or more tissues in a subject in need thereof. In particular, the technology of the present disclosure relates to biological approaches for disrupting serine hydratase (SDS) activity in a subject.

A multiplex assay for nucleic acid detection includes a substrate, a sample, and a fluorophore-labeled oligonucleotide. The substrate has a plurality of physically separated assay locations, each of which includes a nucleotide-targeting enzyme configured to cleave nucleic acids, a guide ribonucleic acid (gRNA), and a quencher-labeled oligonucleotide. A portion of the sample is distributed to each assay location. The gRNA recognizes target nucleic acid in the sample, thereby activating the nucleotide-targeting enzyme to cleave nucleic acids, including the quencher-labeled oligonucleotide. The fluorophore-labeled oligonucleotide is subsequently added to each assay location, which facilitates identification of a presence of the target nucleic acid in the sample via detection of unquenched light emitted by the fluorophore in one or more of the plurality of assay locations.

Described herein is the Phage high-throughput approach for gene essentiality Mapping and Profiling (PhageMaP) platform to generate pooled, barcoded phage knockouts for high throughput screening of conditional phage gene essentiality. A barcoded donor plasmid library is prepared wherein each member includes a single guide RNA (gRNA) spacer sequence and 5 and 3 homology arm sequences. A host cell engineered to express a Cas nuclease and a recombinase is then transformed with the donor plasmid library followed by infection with a population of target phage to provide a barcoded target phage variant library produced by double stranded cleavage of the target phage genomes by the Cas nuclease-gRNA and subsequent recombinase-mediated homologous recombination with the donor plasmid. Insertion of the barcodes at the genomic loci disrupts the function of the genomic loci and provides the barcoded target phage variant library which may then be isolated.

Provided herein are targeting constructs for sustained transgene expression of inducible cell death systems that include mutants of estrogen receptor alpha ligand binding domain (ER-LBD). Also provided are methods for use of the same, such as inducing cell death in a cell.

The present disclosure provides methods for reducing the level of an RNA transcript from a target nucleic acid. The present disclosure provides methods of treating a disease that results from or is caused by a toxic gain-of-function protein. The present disclosure provides systems and compositions for carrying out a method of the present disclosure.