The present disclosure provides methods and systems for DNA modification and gene targeting comprising an engineered Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated transposon (CAST) systems. More particularly, the present disclosure provides systems comprising: an engineered CAST system or one or more nucleic acids encoding the engineered CAST system, wherein the CAST system comprises at least one or both of: a) at least one Cas protein (e.g., Cas6, Cas7, Cas5, and/or Cas8) and b) one or more transposon-associated proteins (e.g., TnsA, TnsB, TnsC, TnsD, and/or TniQ), and at least one unfoldase protein (e.g., ClpX), or a nucleic acid encoding thereof. The present disclosure also provides systems, kits, and methods for nucleic acid modification in a cell.

Provided is use of soybean gene Glyma.13G252100 in regulating resistance of soybean to Phytophthora. The nucleotide sequence of the soybean gene Glyma.13G252100 is set forth in SEQ ID NO: 1. The resistance of the soybean to Phytophthora is enhanced based on a decrease in an expression level of the soybean gene Glyma.13G252100; or the resistance of the soybean to Phytophthora is reduced based on an increase in an expression level of the soybean gene Glyma.13G252100. The present disclosure is the first discovery that modulation of Glyma.13G252100 expression can regulate resistance of soybean to Phytophthora.

Provided herein are compositions and methods for modifying the gene encoding a neonatal fragment crystallizable receptor (FcRn) protein and/or expression or activity thereof in a mammalian cell. The compositions and methods disclosed herein provide variant FcRn proteins having reduced ability to bind to an Fc region of an IgG antibody.

Provided herein are compositions, systems, and methods comprising effector proteins for treating DUX4 mutations. These effector proteins may be characterized as CRISPR-associated (Cas) proteins. Various compositions, systems, and methods of the present disclosure may leverage the activities of these effector proteins for the modification, detection, and engineering the DUX4 gene.

Disclosed are recombinant virions that have a modified capsid protein or a variant thereof of erythroparvovirus and a nucleic acid that includes a heterologous nucleic acid.

Disclosed herein are an engineered immune cell with CIITA gene knock-out and use thereof. According to the present invention, an sgRNA specifically targeting CIITA gene is designed and synthesized, which can accurately target CIITA gene to achieve gene knock-out with high knockout efficiency. The provided sgRNA can be used for preparing a universal CAR-T cell.

Disclosed herein are siRNAs and antisense oligonucleotides (ASOs) specific for an mRNA sequence of a mutS homolog 2 (MSH2) gene, PMS1 homolog 2, mismatch repair system component (PMS2) gene, mutS homolog 6 (MSH6) gene, or mutL homolog 1 (MLH1) gene. Such siRNAs and ASOs can be used in methods of inhibiting DNA mismatch repair. Also disclosed are systems and methods that combine the use of these siRNAs and ASOs with prime editing technology.

Disclosed herein is use of miR528 in production and breeding of gramineous forage grasses. By down-regulating the expression and/or function of miR528, the present invention can significantly enhance the tillering and/or regeneration capacity of the gramineous forage grasses, which is conducive to the biomass accumulation of the gramineous forage grasses, and the breeding of new gramineous forage grass subspecies with enhanced tillering and/or regeneration capacity.

Disclosed are methods for inducing tolerance to Huanglongbing (HLB) in citrus plants. It was found that silencing negative regulators of the plant immune system will provide resistance or tolerance to diseases, particularly HLB. NPR3 is silenced using citrus tristeza virus-based plant mediated RNA interference (CTV-RNAi) to create citrus plants that are resistance to HLB. Also disclosed are CTV-RNAi vectors designed to silence negative regulators of the plant immune system.

EVs are being recognized as vectors for drug delivery. In particular. EV loading with targeting and therapeutic agents brings along an interesting opportunity to translate EVs into a bio-mimetic selective delivery system. Indeed. EVs constitute a physiological carrier being potentially less immunogenic than artificial delivery vehicles. The inventors now developed a novel method to control the loading of a cargo into EVs on demand. These EVs are equipped, if necessary, with non-viral fusogen, therefore enhancing EV-cargo delivery into acceptor cells. To acutely measure this process, they follow the fate of a luciferase-tagged cargo. Cargo loading was enabled through a drug-reversible inducible dimerization system. Briefly, donor cells were transfected with plasmids encoding for FKBP-tagged CD63, a classical membrane EV marker, and FRB-Nanoluciferase (NLuc) that is normally cytosolic. Upon addition of the dimerizing drug. FRB-Nluc interacts with FKBP-CD63 and is recruited into secreted EVs. This is accompanied by an enhanced delivery into acceptor cells. This phenomenon can be further enhanced when EVs are equipped with syncitin1, a mammalian fusogenic protein that trigger fusion between EV membrane and the plasma membrane of acceptor cells. Using this novel process, the inventors further demonstrated that the catalytic domain of the Diphteria toxin (DTA), that is responsible for protein synthesis inhibition and ultimately cell death, can be delivered to acceptor cells via functionalized EVs. This led to protein synthesis inhibition and death of acceptor cells. This novel method and the derived applications promise to open new doors in precision care medicine, especially when EVs will be equipped with antibodies raised against cell specific antigens.