The present invention pertains to extracellular vesicle (EV) therapeutics, wherein the EVs comprise nucleic acid (NA)-based therapeutics such as mRNAs, circular RNAs, miRNAs, shRNAs, circular RNA and/or DNA molecules. The NA therapeutics are loaded into EVs using inventive engineering protein and NA engineering strategies to enhance loading into EVs and to facilitate release of the NA cargo molecules inside target cells.

The present invention relates to retroviral particle comprising a protein derived from the Gag polyprotein, an envelope protein, optionally an integrase and at least two encapsidated non-viral RNAs, the encapsidated non-viral RNAs each comprising an RNA sequence of interest bound to an encapsidation sequence, each encapsidation sequence being recognized by a binding domain introduced into the protein derived from the Gag polyprotein and/or into the integrase, and at least one of said sequences of interest of the encapsidated non-viral RNAs comprises a part coding at least one epitope and/or at least one molecular structure specifically recognizing an epitope.

Disclosed are RNA site-directed editing using artificially constructed RNA editing enzymes and related uses. Provided is the fusion of an RNA recognition domain for binding RNA and a functional effector domain to form a new functional protein. The new functional protein specifically targets target RNA by means of the recognition domain and performs RNA editing using the effector domain.

A system for editing of a target sequence at a locus of a host cell is disclosed. The system has a nucleic acid molecule comprising a nucleic acid segment comprising a targeting RNA sequence and an RNA segment that binds a protein. The system also has a nucleic acid molecule comprising a nucleic acid segment encoding a polypeptide with endonuclease activity fused to a protein that binds the RNA segment. The system also comprises a double stranded DNA molecule comprising DNA comprising at least one nucleotide sequence that is capable of binding to the target sequence at the locus.

The present invention belongs to the field of plant genetic engineering. Specifically, the invention relates to a method for creating novel herbicide resistant plants by base editing techniques and a method for screening endogenous gene mutation sites capable of conferring herbicide resistance in plants. The invention also relates to the use of the identified endogenous gene mutantation sites in crop breeding.

Disclosed herein are nucleic acid-guided nucleases, guide nucleic acids, and targetable nuclease systems, and methods of use. Disclosed herein are engineered non-naturally occurring nucleic acid-guided nucleases, guide nucleic acids, and targetable nuclease systems, and methods of use. Targetable nuclease systems can be used to edit genetic targets, including recursive genetic engineering and trackable genetic engineering methods.

A method for designing a protein capable of binding in an RNA base selective manner or RNA base sequence specific manner is provided. The protein of the present invention is a protein containing one or more of PPR motifs (preferably 2 to 14 PPR motifs) each consisting of a polypeptide of 30- to 38-amino acid length represented by the formula 1 (wherein Helix A is a moiety of 12-amino acid length capable of forming an α-helix structure, and is represented by the formula 2, wherein, in the formula 2, A.sub.1 to A.sub.12 independently represent an amino acid; X does not exist, or is a moiety of 1- to 9-amino acid length; Helix B is a moiety of 11- to 13-amino acid length capable of forming an α-helix structure; and L is a moiety of 2- to 7-amino acid length represented by the formula 3, wherein, in the formula 3, the amino acids are numbered “i” (−1), “ii” (−2), and so on from the C-terminus side, provided that L.sub.iii to L.sub.vii may not exist), and combination of three amino acids A.sub.1, A.sub.4 and L.sub.ii, or combination of two amino acids A.sub.4, and L.sub.ii is a combination corresponding to a target RNA base or base sequence.

Disclosed is a method and system for the phase separation of folded domains, and more particularly, to inducing clusters of folded domains as part of a drug-based screening application. The system and method utilize one or more first fusion proteins (100, 101), each first fusion protein comprising a first region (110) fused to a second region (120), the first region (110) comprising at least one light sensitive protein (115) or cognate partner of a light sensitive protein (116), and the second region (120) comprising one or more folded RNA binding domains (RBDs), disordered RBDs, folded non-RBD domains, or combination thereof (125).

The invention relates to a method of altering a differentiation status of a stem cell by modulating the expression of one or more differentiation factors with a nuclease-deactivated Cas9 (dCas9) fusion protein comprising dCas9 and a transcriptional activator. The method may further include a guide RNA (gRNA) and an activator module comprising RNA-binding protein binding capable of binding to the gRNA. In one embodiment, the dCas9 fusion protein comprises dCas9 and VP64 while the activator module comprises MS2 coat protein and p65. The one or more differentiation factors may comprise PAX6, MITF and OTX2 for differentiation of pluripotent stem cell into retinal pigment epithelium (RPE). Also disclosed are cells comprising the dCas9 fusion protein, gRNA, kits, and method of treating a disease thereof.

This disclosure relates to Cas9-nucleic acid complexes and uses related thereto. In certain embodiments, the disclosure contemplates transgenic plants and animals genetically engineered to express Cas9-nucleic acid complexes disclosed herein. In certain embodiments, the disclosure relates to methods of treating or preventing, diseases, conditions, cancer, viral infections or other pathogenic infection using vectors configured to express a Cas9-nucleic acid complex disclosed herein.