The present disclosure provides polynucleotides and vectors for the genomic engineering and editing of non-conventional yeasts such as Issatchenkia orientalis. The polynucleotides and vectors can be used as tools that are efficient to alter the expression of one or more gene products in the yeast, and specifically to induce the production of organic acids or other bioproducts of interest in the yeast.

Disclosed are components and methods for producing toxic RNAs and/or extracellular vesicles comprising the toxic RNAs in cells that are resistant to the toxic RNAs, which toxic RNAs may include toxic shRNA/siRNA and/or toxic pre-miRNA/miRNA. The disclosed components may include engineered cells that can be utilized to express the toxic RNAs, in which the engineered cells do not express one or more genes that are required for processing the toxic RNAs for RNA interference (RNAi) and/or one or more genes that are required for executing RNAi. The toxic RNAs and/or extracellular vesicles comprising the toxic RNAs may be utilized in methods for treating diseases and disorders through RNAi.

Compositions for improved gene editing and methods of use thereof are disclosed. In a preferred method, gene editing involves use of a cell-penetrating anti-DNA antibody, such as 3E10, as a potentiating agent to enhance gene editing by nucleases and triplex forming oligonucleotides. Genomic modification occurs at a higher frequency when cells are contacted with the potentiating agent and nuclease or triplex forming oligonucleotide, as compared to the absence of the potentiating agent. The methods are suitable for both ex vivo and in vivo approaches to gene editing and are useful for treating a subject with a genetic disease or disorder. Nanoparticle compositions for intracellular delivery of the gene editing compositions are provided and are particularly advantageous for use with in vivo applications.

Compositions for enhanced gene editing and methods of use thereof are. The composition contains a cell-penetrating antibody and a donor oligonucleotide containing a sequence that can correct a mutation in a cell's genome. Preferably, the composition does not contain a nuclease, PNA, or nanoparticle. The compositions are used to modify the genome of a cell by contacting the cell with an effective amount of the composition. Genomic modification occurs at a higher frequency both ex vivo and in vivo, when cells are contacted with the cell-penetrating antibody and donor oligonucleotide as compared to the absence of the cell-penetrating antibody.

The subject invention provides methods, assays, and products for detecting small-molecule targets in a complex sample in both clinical and field settings. The subject invention provides aptamer-based sensors and methods of use thereof. The subject invention provides exonuclease-based methods for generating structure-switching aptamers from fully folded or pre-folded aptamers and developing aptamer-based sensors for small-molecule detection. The method for detecting one or more small-molecule targets in a sample comprises contacting the sample with one or more aptamer-based sensor selective for each of the small-molecule targets, and detecting the small-molecule target in the sample.

Provided is an α-amylase which can function at low temperatures while maintaining or enhancing stability and/or cleaning performance. An α-amylase mutant, which is a mutant of a parent α-amylase, the α-amylase mutant comprising one or more modifications to amino acid residues at positions corresponding to positions G5, S38, T49, Q96, N126, T129, G140, F153, Q167, G179, W186, E187, N192, M199, Y200, L203, Y205, D206, R211, K215, H240, S241, Y242, G244, E257, F259, K278, H283, S284, A288, H295, Y296, N303, T320, S331, L348, Y360, W408, L429, V430, G433, A434, W439, N471, G476, and G477 in the amino acid sequence of SEQ ID NO: 2, the parent α-amylase or α-amylase mutant having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 4.

Compositions for improved gene editing and methods of use thereof are disclosed. In a preferred method, gene editing involves use of a cell-penetrating anti-DNA antibody, such as 3E10, as a potentiating agent to enhance gene editing by nucleases and triplex forming oligonucleotides. Genomic modification occurs at a higher frequency when cells are contacted with the potentiating agent and nuclease or triplex forming oligonucleotide, as compared to the absence of the potentiating agent. The methods are suitable for both ex vivo and in vivo approaches to gene editing and are useful for treating a subject with a genetic disease or disorder. Nanoparticle compositions for intracellular delivery of the gene editing compositions are provided and are particularly advantageous for use with in vivo applications.

The present disclosure provides a method of treating or preventing metabolic disorders by administering agents that inhibit the activity of microRNAs that modulate metabolism.

Methods of modulating expression of a target nucleic acid in a cell are provided including use of multiple orthogonal Cas9 proteins to simultaneously and independently regulate corresponding genes or simultaneously and independently edit corresponding genes.

The present disclosure describes compositions of matter comprising a ribonucleoprotein complex comprising a nucleic acid-guided nuclease and a guide RNA, and further comprising and a blocking nucleic acid molecule represented by Formula I, wherein Formula I in the 5′-to-3′ direction comprises: A-(B-L).sub.J-C-M-T-D; wherein A is 0-15 nucleotides in length; B is 4-12 nucleotides in length; L is 3-25 nucleotides in length; J is an integer between 1 and 10; C is 4-15 nucleotides in length; M is 1-25 nucleotides in length or is absent, wherein if M is absent then A-(B-L).sub.J-C and T-D are separate nucleic acid strands; T is 17-135 nucleotides in length and comprises at least 50% sequence complementarity to B and C; D is 0-10 nucleotides in length and comprises at least 50% sequence complementarity to A; and wherein the blocking nucleic acid molecule comprises a sequence complementary to a gRNA.