This invention relates generally to segmented oligonucleotides capable of modulating gene expression. Specifically, the instant invention relates to segmented microRNA (miRNA) oligonucleotides, including segmented miRNA precursors and segmented pre-microRNAs. The invention also relates to compositions comprising such segmented oligonucleotides, as well as to methods of making and using such oligonucleotides for diagnosis and treatment of diseases associated or causally linked to aberrant levels or activities of gene expression, including aberrant levels of coding and/or non-coding RNA.

The present disclosure discloses use of a miRNA 148 cluster as a marker for diagnosing and/or treating cognitive impairment-associated diseases. The present disclosure provides use of the miRNA 148 cluster in the diagnosis and treatment of cognitive impairment-associated diseases. The expression level of the miRNA 148 cluster is detected using primers for the microRNA through cognitive impairment-associated disease models, and it is found that the expression of the miRNA 148 cluster is significantly reduced during the progression of the cognitive impairment-associated diseases. The miRNA 148 cluster is found to directly target p35 to inhibit hyperphosphorylation of Tau and can be negatively regulated by the upregulated PTEN in AD pathology. Upregulation of miRNA 148 cluster improves the cognitive dysfunction and inhibit the hyperphosphorylation of Tau in AD pathology, in which the PTEN/Akt/CREB and p35/CDK signaling pathways play a key role.

The present disclosure provides DNA-guided CRISPR systems; polynucleotides comprising DNA, RNA and mixtures thereof for use with CRISPR systems; and methods of use involving such polynucleotides and DNA-guided CRISPR systems.

Novel oligonucleotide pairs which can form a duplex comprising one or more DNA-like nucleotides (e.g., 2′-substituted arabinonucleotides (ANA)); in combination with one or more RNA-like nucleotides (e.g., 2′-substituted ribonucleotides (RNA) and/or locked nucleic acid nucleotides (LNA)), are disclosed. The use of such oligonucleotide duplexes, such as for silencing the expression of a nucleic acid or gene of interest using small interfering RNA (siRNA) technologies, is also disclosed.

The present invention concerns methods and reagents useful in modulating gene expression in a variety of applications, including use in therapeutic, diagnostic, target validation, and genomic discovery applications. Specifically, the invention relates to synthetic chemically modified small nucleic acid molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA) molecules capable of mediating RNA interference (RNAi) against target nucleic acid sequences. The small nucleic acid molecules are useful in the treatment of any disease or condition that responds to modulation of gene expression or activity in a cell, tissue, or organism.

The present invention provides stable, nuclease-resistant TNA and TNA-DNA oligonucleotides, wherein the oligonucleotides are completely resistant to enzymatic degradation for at least 24-72 hours. Methods of synthesis and use in diagnostic and therapeutic applications are also provided. Specifically, in one embodiment, we describe the chemical and biological stability of TNA and mixed-backbone (mosaic) TNA-DNA oligonucleotides under a variety of conditions and sequence contexts.

Disclosed is a modified oligonucleotide capable of reducing or inhibiting one or more activities miR-138. The modified oligonucleotide may comprise at least one locked nucleic acid (LNA) and wherein the oligonucleotide is substantially complementary to a nucleotide sequence of miR-138. Also disclosed are pharmaceutical compositions comprising the oligonucleotides, methods of using the oligonucleotides and uses thereof.

Nucleic acids (DNA and RNA) provide a versatile platform for engineering synthetic biology in a variety of technology areas including medicine, science, agriculture, and energy. In many settings, degradation of nucleic acid molecules poses a significant engineering challenge as the molecules do not function if they have been degraded. In some embodiments, nucleic acid protective elements (PELs) are used to protect chemically synthesized or expressed nucleic acid molecules from degradation. PELs may be derived from all or part of a viral xrRNA sequence and/or structural motif, PELs may include rationally designed sequences and/or structural motifs, PELs may be engineered using directed evolution, and in some embodiments, PELs comprise a mixture of biologically derived, rationally designed sequence and/or structural motifs, and/or sequences and/or structural motifs that are engineered by directed evolution. In some embodiments, PELs significantly enhance the performance of nucleic acid synthetic biology, protecting nucleic acid regulatory and/or structural elements from degradation to increase regulatory dynamic range, fractional dynamic range, fold-change, and/or other performance metrics. In some embodiments, PELs that reduce nucleic acid degradation provide a platform technology for enhancing the performance of synthetic biology, with applications including therapeutics, diagnostics, biological research tools, vaccines, crop protection, molecular manufacturing, sustainable energy production, and other areas involving nucleic acids.

Neutral lipid formulations for nucleic acid delivery are provided according to the invention. The neutral lipid formulations include hydrophobically modified polynucleotides and fat mixtures. Methods of using the neutral lipid formulations are also provided.

The present invention relates to RNAi agents, e.g., double stranded RNA (dsRNA) agents, targeting the SERPINF2 gene. The invention also relates to methods of using such RNAi agents to inhibit expression of a SERPINF2 gene and to methods of preventing and treating a SERPINF2-associated disorder, e.g., a disorder associated with thrombosis.