Wisconsin Alumni Research Foundation

Amr Ashraf Abdeen

Krishanu Saha

Shaoqin Gong

Yuyuan Wang

Guojun Chen

Jared Matthew Carlson-Stevermer

Lucille Katherine Kohlenberg

Ruosen Xie

Provided herein are nanoplexes comprising a payload selected from a protein and/or a polynucleic acid; and a plurality of copolymers comprising a first copolymer that is poly(N,N'-bis(acryloyl)cystamine-poly(aminoalkyl)) (PBAP), a second copolymer that is poly(Cakylene glycol)-PBAP-poly(Cakylene glycol), and a third copolymer that is TG-poly(Cakylene glycol)-PBAP-poly(Cakylene glycol)-TG wherein TG at each occurrence is independently a targeting ligand, a cell penetrating peptide, an imaging agent or a capping group, provided that a plurality of TG groups is a targeting ligand; wherein the payload is non-covalently complexed to one or more of the copolymers, one or more of the first, second, and/or third copolymers comprises an endosomal escape group having a pKa of about 4.5 to about 6.5, and optionally one or more of the first, second, and/or third copolymers comprises a host and a guest non-covalent crosslinker.

Polyplex delivery system for proteins, nucleic acids and protein/nucleic acid complexes

Provided herein are nanocapsules comprising a single ribonucleoprotein (RNP) complex as a core and an biodegradable crosslinked polymer shell that encapsulates the core, wherein the RNP complex comprises a Cas9 polypeptide and a guide RNA, and the biodegradable crosslinked polymer shell comprises polymerized monomers of imidazolyl acryloyl monomers, bisacryloyl disulfide monomers (a biodegradable cross-linker), optionally PEG acryloyl monomers, and either cationic acryloyl monomers, anionic acryloyl monomers, or both cationic and anionic acryloyl monomers (optionally in combination with non-ionic acryloyl monomers) as defined herein. Also provided are methods of making the nanocapsules, kits containing the nanocapsules and methods of delivering the encapsulated RNP to cells.

Nanocapsule delivery system for ribonucleoproteins

Described herein are modified guide RNAs such as a single guide RNA including, from 5' to 3&#x2032;, a single-stranded protospacer sequence, a first complementary strand of a binding region for the Cas9 polypeptide, an aptamer that binds a biotin-binding molecule, and a second complementary strand of the binding region for the Cas9 polypeptide. Also described is an RNP complex including the modified guide RNA and a Cas9 polypeptide or active fragment thereof. Also included are methods of modifying target genes in cells using the modified guide RNAs.

Modified guide RNAs, CRISPR-ribonucleotprotein complexes and methods of use

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Madison, WI, United States of America

Amr Ashraf Abdeen