University of Massachusetts

Massachusetts Institute of Technology

Max-planck-gesellschaft Zur F&#xf6;rderung Der Wissenschaften E. V.

Whitehead Institute for Biomedical Research

Juanita Mclachlan

Phillip D. Zamore

Craig C. Mello

Alla Grishok

Gyorgy Hutvagner

Phillip D Zamore

Guangping Gao

Dianne S Schwarz

Thomas Tuschl

Phillip A Sharp

David P Bartel

Craig C Mello

Juanita McLachlan

Neil Aronin

Guiliang Tang

Martin Simard

Jun Xie

Christian Mueller

Terence Flotte

György Hutvágner

Yi Lu

Dan Wang

Phillip Tai

Jennifer Broderick

Benjamin Haley

Qiang Zheng

Xun Xu

Edith L Pfister

Melissa Moore

David Grunwald

Maximiliaan Huisman

Carlas Smith

Shaoyong Li

Karina Jouravleva

Christian Roy

Xu Xun

Gyoergy Hutvagner

Dianne Schwarz

Guanping Gao

György HUTVÁGNER

Gyorgy Hutvanger

The present disclosure relates to the field of rAAV delivery of transgenes. In some aspects, the disclosure relates to RNAi. Provided herein are recombinant adeno-associated virus (rAAV) vectors comprising modified ITRs. In some embodiments, the modified ITRs comprise a sequence encoding a shRNA, miRNA, or AmiRNA.

Modified AAV constructs and uses thereof

The disclosure in some aspects relates to recombinant adeno-associated viruses having nuclease grafted to one or more capsid proteins. In some aspects, the disclosure relates to isolated AA V capsid proteins having terminally grafted nucleases and isolated nucleic acids encoding the same. Recent approaches to delivering nucleases to cells for gene editing have focused on delivering of expression vectors engineered to express the nucleases in target cells. However, these approaches have proved to be problematic in many instances due to genotoxicity resulting from to prolonged expression of gene editing system in vivo. To prevent such off-target genotoxicity due to prolonged presence of a gene editing system, several studies explored delivery of mRNA or protein instead of delivering the gene coding for the nucleases in cell culture.

Compositions and methods for transient delivery of nucleases

The invention relates to isolated nucleic acids and rAAV-based compositions, methods and kits useful for treating genetic diseases (e.g., alpha-1 antitrypsin deficiency).

RAAV-based compositions and methods

The disclosure relates, in some aspects, to compositions and methods for treating corneal disease (e.g., corneal neovascularization. In some embodiments, the disclosure relates to rAAV-mediated delivery of an cornea-associated transgene to a subject. In some embodiments, the rAAV transduces the corneal tissue of a subject.

Anti-angiogenic miRNA therapeutics for inhibiting corneal neovascularization

The invention is directed to a heated stage assembly that can achieve high temperatures (i.e., 40 degrees Celsius and higher and more specifically, 55 degrees Celsius and higher) for use in microscopy including fluorescence microscopy. The microscope stage heater assembly includes a mounting adapter element, a sample holder element mounted within the mounting adapter element, one or more heating elements attached to the sample holder element, and an optical window holder element disposed on a surface of the sample holder element that encloses an optical window element. The optical window element may be in contact with a microscope sample such that the optical window element is heated by the sample holder element to uniformly distribute heat to the specimen.

Heated stage assembly for high temperature fluorescence microscopy

The invention in some aspects relates to isolated nucleic acids, compositions, and kits useful for identifying adeno-associated viruses in cells. In some aspects, the invention provides kits and methods for producing somatic transgenic animal models using recombinant AAV (rAAV) to an animal having at least one transgene that expresses a small interfering nucleic acid or at least one binding site for a miRNA.

Isolation of novel AAV's and uses thereof

The instant disclosure provides RNA-modulating agents that function to recruit one or more small regulatory RNA molecules (e.g., miRNA molecules, Y RNAs, and siRNAs) to a target mRNA thereby modulating (e.g., inhibiting) the translation of the target mRNA or destabilizing the mRNA. Also provided are miRNA inhibitors and diagnostic agents that have improved binding affinity for their target miRNAs. Methods for using the RNA-modulating agents, miRNA inhibitors and diagnostic agents are also provided.

RNA-modulating agents

Based at least in part on an understanding of the mechanisms by which small RNAs (e.g., naturally-occurring miRNAs) mediate RNA silencing in plants, rules have been established for determining, for example, the degree of complementarity required between an RNAi-mediating agent and its target, i.e., whether mismatches are tolerated, the number of mismatches tolerated, the effect of the position of the mismatches, etc. Such rules are useful, in particular, in the design of improved RNAi-mediating agents which allow for more exact control of the efficacy of RNA silencing.

Methods and compositions for controlling efficacy of RNA silencing

The invention in some aspects relates to methods and compositions for assessing the effectiveness of miRNA inhibitors. In other aspects of the invention, methods and compositions for treating cholesterol related disorders are provided. In one aspect of the invention, miRNA inhibitors against miR-122 and rAAV-based compositions comprising the same are provided.

AAV-based treatment of cholesterol-related disorders

The present invention relates to the discovery of a method for inhibiting RNA silencing in a target sequence-specific manner. RNA silencing requires a set of conserved cellular factors to suppress expression of gene-encoded polypeptide. The invention provides compositions for sequence-specific inactivation of the RISC component of the RNA silencing pathway, and methods of use thereof. The RISC inactivators of the present invention enable a variety of methods for identifying and characterizing miRNAs and siRNAs, RISC-associated factors, and agents capable of modulating RNA silencing. Therapeutic methods and compositions incorporating RISC inactivators and therapeutic agents identified through use of RISC inactivators are also featured.

Sequence-specific inhibition of small RNA function

The present invention relates to the discovery of an effective treatment for a variety of gain-of-function diseases, in particular, Huntington's disease (HD). The present invention utilizes RNA Interference technology (RNAi) against polymorphic regions in the genes encoding various gain-of-function mutant proteins resulting in an effective treatment for the gain-of-function disease.

RNA interference for the treatment of gain-of-function disorders

The disclosure in some aspects, relates to nucleic acids, compositions and kits useful for gene therapy with reduced immune response to transgene products.

Recombinant AAV vectors useful for reducing immunity against transgene products

Growing community of inventors

Northboro, MA, United States of America

Phillip D Zamore