A very fast and efficient CRISPR/Cas9 system is provided. Compositions include light-sensitive caged nucleotides at the PAM distal region of guide RNAs (gRNA.sup.caged) to create artificial mismatches as a “roadblock”. Upon light stimulation, the caging group (“roadblock”) is removed and the gRNA fully hybridizes with the target DNA. Thus, the pre-bound inactive Cas9/gRNA.sup.caged is rapidly converted to active Cas9.

The invention provides systems and methods for discovering candidate therapies for genetic conditions and also for screening those therapies in vitro for evidence of neurotoxicity. Where a medical condition is a consequence of a genetic target such as a mutated gene, the disclosure provides in silico methods to generate lists of candidate sequences for antisense oligonucleotides (ASOs) that will potentially bind to the gene or transcripts from the gene in vivo and treat the associated condition by restoring a healthy phenotype of gene expression. The invention provides in vitro methods for screening candidate ASO sequences for symptoms of neurotoxicity in vivo. For example, candidate sequences that are output by the in silico analytical pipeline can be synthesized and assayed against live cells in vitro.

The present invention provides a method for producing a nucleic acid molecule that can obtain a nucleic acid molecule that binds to a target and does not inhibit a function of the target. The production method for a nucleic acid molecule of the present invention is a method for producing a nucleic acid molecule that binds to a first biological molecule and does not inhibit a function of the first biological molecule, the method including the steps of: (A) bringing a candidate nucleic acid molecule into contact with the first biological molecule to select a nucleic acid molecule that has bound to the first biological molecule as a first selected nucleic acid molecule; and (B) selecting the first selected nucleic acid molecule as an intended nucleic acid molecule.

The present invention relates to a method for providing information on the diagnosis of Parkinson's disease. The present invention also relates to a composition for preventing, ameliorating or treating Parkinson's disease. The present invention uses at least one miRNA whose expression is specifically down- or up-regulated in a Parkinson's disease model. Therefore, the use of the miRNA is effective in diagnosing and treating Parkinson's disease.

Non-human animal cells and non-human animals comprising CRISPR/Cas synergistic activation mediator system components and methods of making and using such non-human animal cells and non-human animals are provided. Methods are provided for using such non-human animals to increase expression of target genes in vivo and to assess CRISPR/Cas synergistic activation mediator systems for the ability to increase expression of target genes in vivo.

Disclosed herein are inhibitors of PPM 1 A, including PPM1A antisense oligonucleotide sequences, and methods for treating neurological diseases, such as amyotrophic lateral sclerosis and frontotemporal dementia, associated with decreased activity or expression of TBK1. Also disclosed are pharmaceutical compositions containing a PPM1A inhibitor, including a PPM1A antisense oligonucleotide, useful for treating neurological diseases and manufacture of medicaments containing a disclosed PPM1A inhibitor, for example, a PPM1A antisense oligonucleotide, to be used in treating a neurological disease

The disclosure relates to immune cells for use in adoptive cell therapy that express an inhibitory receptor, useful for treating a disease or disorder, for example, cancer. The disclosure provides immune cells with reduced or eliminated HLA expression, that express an inhibitory receptor, methods of making same, shRNAs targeting HLA-A mRNA, and polynucleotides and vectors encoding same.

This disclosure relates to oligonucleotides, compositions and methods useful for reducing GYS2 expression, particularly in hepatocytes. Disclosed oligonucleotides for the reduction of GYS2 expression may be double-stranded or single-stranded, and may be modified for improved characteristics such as stronger resistance to nucleases and lower immunogenicity. Disclosed oligonucleotides for the reduction of GYS2 expression may also include targeting ligands to target a particular cell or organ, such as the hepatocytes of the liver, and may be used to treat glycogen storage diseases (e.g., GSDIa, GSDIII, GSDIV, GSDVI, and GSDIX) and related conditions.

RNAs, such as miRNA and siRNA, and their use in treating complement-mediated disorders, are described.

The invention provides for delivery, engineering and optimization of systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are delivery systems and tissues or organ which are targeted as sites for delivery. Also provided are vectors and vector systems some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells to ensure enhanced specificity for target recognition and avoidance of toxicity and to edit or modify a target site in a genomic locus of interest to alter or improve the status of a disease or a condition.