This application relates to therapeutic siRNA agents and methods of making and using the agents.

The present invention relates to the field of structural biology. More specifically, the present invention relates to an antigen-binding chimeric protein, called a MegaBody™, specifically binding a nanodisc, more specifically a membrane-scaffold protein (MSP)n which may be part of the nanodisc. The invention further provides for methods and uses of said nanodisc-specific antigen-binding chimeric proteins in three-dimensional high-resolution structural analysis of membrane proteins assembled within nanodiscs. The MSP-binding MegaBodies of the invention provide for a generic tool in membrane protein structural biology, more particular in Cryo-EM, by reducing preferred particle orientation of nanodiscs and of the entrapped target membrane proteins.

The present invention relates to the field of structural biology. More specifically, the present invention relates to an antigen-binding chimeric protein, called a MegaBody™, specifically binding a nanodisc, more specifically a membrane-scaffold protein (MSP)n which may be part of the nanodisc. The invention further provides for methods and uses of said nanodisc-specific antigen-binding chimeric proteins in three-dimensional high-resolution structural analysis of membrane proteins assembled within nanodiscs. The MSP-binding MegaBodies of the invention provide for a generic tool in membrane protein structural biology, more particular in Cryo-EM, by reducing preferred particle orientation of nanodiscs and of the entrapped target membrane proteins.

The present disclosure features methods and compositions for treating Alzheimer's disease. The disclosed methods comprise administering to a subject having or suspected of having Alzheimer's a hematopoietic stem progenitor cell expressing at least one neuroprotective agent, such as ApoE2, Trem2, and/or a metallothionein.

The present disclosure relates to the field of biotechnology, in particular to a base editing tool and use thereof. The present disclosure provides a fusion protein comprising a first nCas9 fragment, a chimeric insertion fragment, a second nCas9 fragment and two UGI fragments from N-terminus to C-terminus, wherein the chimeric insertion fragment is selected from APOBEC1 fragment or APOBEC3A fragment. The present disclosure provides a novel base editing tool that is compatible with insertion of various deaminases on the chimeric sites of nCas9. Compared with nCas9 terminal fusion base editor, the base editing tool of the present invention significantly reduce of off-targeting on both DNA and RNA, while maintaining specific targeted base editing efficiency, with higher specificity and favorable industrialization prospects.

The present disclosure relates to the field of biotechnology, in particular to a base editing tool and use thereof. The present disclosure provides a fusion protein comprising a first nCas9 fragment, a chimeric insertion fragment, a second nCas9 fragment and two UGI fragments from N-terminus to C-terminus, wherein the chimeric insertion fragment is selected from APOBEC1 fragment or APOBEC3A fragment. The present disclosure provides a novel base editing tool that is compatible with insertion of various deaminases on the chimeric sites of nCas9. Compared with nCas9 terminal fusion base editor, the base editing tool of the present invention significantly reduce of off-targeting on both DNA and RNA, while maintaining specific targeted base editing efficiency, with higher specificity and favorable industrialization prospects.

The invention provides expression cassettes. In certain embodiments, an expression cassette comprises (a) a regulatory element at least 90% identical to the sequence of any of SEQ ID NOs:2-67, and (b) a nucleic acid sequence encoding a Factor VIII protein having a B domain deletion (FVIII-BDD), where the nucleic acid sequence of (a) is at least 90% identical to the sequence of SEQ ID NO:77, where the regulatory element is operably linked to the nucleic acid sequence, and where no intron is present between the regulatory element and the nucleic acid sequence encoding FVIII-BDD, or where no more than 0-107 nucleotides of untranslated nucleic acid is between the regulatory element and the nucleic acid sequence encoding FVIII-BDD. In certain embodiments, expression cassettes contain sequence elements having CpG(s) substituted with CpT, CpA, TpG, or ApG at the same position(s) or has CpG reduced nucleic acid sequences.

The invention provides expression cassettes. In certain embodiments, an expression cassette comprises (a) a regulatory element at least 90% identical to the sequence of any of SEQ ID NOs:2-67, and (b) a nucleic acid sequence encoding a Factor VIII protein having a B domain deletion (FVIII-BDD), where the nucleic acid sequence of (a) is at least 90% identical to the sequence of SEQ ID NO:77, where the regulatory element is operably linked to the nucleic acid sequence, and where no intron is present between the regulatory element and the nucleic acid sequence encoding FVIII-BDD, or where no more than 0-107 nucleotides of untranslated nucleic acid is between the regulatory element and the nucleic acid sequence encoding FVIII-BDD. In certain embodiments, expression cassettes contain sequence elements having CpG(s) substituted with CpT, CpA, TpG, or ApG at the same position(s) or has CpG reduced nucleic acid sequences.

A method for reducing neuronal and synaptic degeneration or loss in a population of neuronal cells is provided as well as a method of treating an individual with a neurocognitive disorder. Aspects of the methods include modulating the level and/or activity of apolipoprotein E (apoE) in a population of neuronal cells where the modulating reduces the level and/or activity of an MHC pathway polypeptide in the population of neuronal cells.

A method for reducing neuronal and synaptic degeneration or loss in a population of neuronal cells is provided as well as a method of treating an individual with a neurocognitive disorder. Aspects of the methods include modulating the level and/or activity of apolipoprotein E (apoE) in a population of neuronal cells where the modulating reduces the level and/or activity of an MHC pathway polypeptide in the population of neuronal cells.