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公开(公告)号：US20160017368A1
公开(公告)日：2016-01-21
申请号：US14871327
申请日：2015-09-30
Applicant: Wisconsin Alumni Research Foundation
Inventor： William L. Murphy , Andrew Salim Khalil , Xiaohua Yu , Krishanu Saha , Jared Matthew Carlson-Stevermeyer , Benjamin Gabriel Steyer
IPC: C12N15/85
Abstract: Disclosed are methods for cell transfection and regulating cellular behavior. More particularly, the present disclosure relates to methods of non-viral cell transfection and regulating cellular behavior using mineral coatings that allow for the enhanced transfection of cells with reduced cytotoxicity. The mineral coatings bind biomaterials and provide a source of calcium and phosphate ions to enhance transfection. The present disclosure also provides a high throughput platform for screening non-viral transfection of cells. The methods of the present disclosure also provide an advantageous biomaterial delivery platform because the mineral coatings may be deposited on various medical device materials after being specifically developed using the high throughput screening platform.
Abstract translation: 公开了细胞转染和调节细胞行为的方法。 更具体地说，本公开内容涉及非病毒细胞转染的方法，并且使用矿物包衣调节细胞行为，其允许增强细胞毒性转染细胞。 矿物涂层结合生物材料，并提供钙和磷酸根离子的来源，以增强转染。 本公开还提供用于筛选细胞的非病毒转染的高通量平台。 本公开的方法还提供有利的生物材料递送平台，因为矿物涂层可以在使用高通量筛选平台特别开发之后沉积在各种医疗装置材料上。
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2.发明授权公开(公告)号：US11739320B2
公开(公告)日：2023-08-29
申请号：US16674448
申请日：2019-11-05
Applicant: Wisconsin Alumni Research Foundation
IPC: C12N15/11 , C12N9/22 , A61P3/00 , A61K35/545 , C12N15/86 , A61K31/7088
CPC classification number: C12N15/11 , A61K31/7088 , A61K35/545 , A61P3/00 , C12N9/22 , C12N15/86 , C12N2310/20 , C12N2310/351 , C12N2800/80
Abstract: Described herein are guide RNAs and modified guide RNAs suitable for biallelic correction of Pompe disease. Also included are methods of modifying a target gene in a patient or in a patient-derived cell, wherein the patient has an autosomal recessive disorder with compound heterozygous mutations, the methods including delivering a first modified guide RNA, a second modified guide RNA, a Cas9 polypeptide, a biotin-binding molecule, a first biotinylated donor polynucleotide, and a second biotinylated donor polynucleotide. The first modified guide RNA and the first biotinylated donor polynucleotide correct a first diseased allele, and the second modified guide RNA and the second biotinylated donor polynucleotide correct a second diseased allele.
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公开(公告)号：US11013695B2
公开(公告)日：2021-05-25
申请号：US16113963
申请日：2018-08-27
Applicant: Wisconsin Alumni Research Foundation
Inventor： Shaoqin Gong , Amr Abdeen , Krishanu Saha , Guojun Chen , Yuyuan Wang , Ruosen Xie
IPC: A61K9/51 , A61K48/00 , A61K47/32 , C12N15/11 , A61K38/46 , C12N15/113 , A61K31/7105 , A61K9/50 , C12N9/22 , C12N15/90
Abstract: 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.
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公开(公告)号：US20240226152A9
公开(公告)日：2024-07-11
申请号：US18295036
申请日：2023-04-03
Applicant: Wisconsin Alumni Research Foundation
Inventor： Krishanu Saha , Lauren Sarko
IPC: A61K35/17 , A61K39/00 , C07K14/005 , C07K16/28 , C12N5/00 , C12N5/0783 , C12N9/22 , C12N15/11
CPC classification number: A61K35/17 , A61K39/4611 , A61K39/4631 , A61K39/464429 , C07K14/005 , C07K16/2896 , C12N5/0056 , C12N5/0636 , C12N9/22 , C12N15/11 , A61K2239/15 , A61K2239/17 , A61K2239/21 , A61K2239/22 , C07K2317/622 , C12N2310/10
Abstract: Described herein are methods using CRISPR-Cas9 and DNA templates that can generate chimeric antigen receptors (CARs) on T cells to target the cell surface protein urokinase Plasminogen Activator Receptor (uPAR) on senescent cells. Also described are methods of preparing CAR T cells, their use to treat neurodegenerative disease, stroke, craniocerebral trauma and/or accident, or elderly individuals in need of treatment for aging.
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公开(公告)号：US11351263B2
公开(公告)日：2022-06-07
申请号：US16282174
申请日：2019-02-21
Applicant: Wisconsin Alumni Research Foundation
Inventor： Shaoqin Gong , Yuyuan Wang , Krishanu Saha , Amr Ashraf Abdeen
Abstract: 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(C2-3 akylene glycol)-PBAP-poly(C2-3 akylene glycol), and a third copolymer that is TG-poly(C2-3 akylene glycol)-PBAP-poly(C2-3 akylene 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.
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Patent Agency Ranking
公开(公告)号：US20180362971A1
公开(公告)日：2018-12-20
申请号：US16008376
申请日：2018-06-14
Applicant: WISCONSIN ALUMNI RESEARCH FOUNDATION
Inventor： Jared Matthew Carlson-Stevermer , Krishanu Saha , Amr Ashraf Abdeen , Lucille Katherine Kohlenberg
Abstract: Described herein are modified guide RNAs such as a single guide RNA including, from 5′ to 3′, 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.
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公开(公告)号：US20240131066A1
公开(公告)日：2024-04-25
申请号：US18295036
申请日：2023-04-02
Applicant: Wisconsin Alumni Research Foundation
Inventor： Krishanu Saha , Lauren Sarko
IPC: A61K35/17 , A61K39/00 , C07K14/005 , C07K16/28 , C12N5/00 , C12N5/0783 , C12N9/22 , C12N15/11
CPC classification number: A61K35/17 , A61K39/4611 , A61K39/4631 , A61K39/464429 , C07K14/005 , C07K16/2896 , C12N5/0056 , C12N5/0636 , C12N9/22 , C12N15/11 , A61K2239/15 , A61K2239/17 , A61K2239/21 , A61K2239/22 , C07K2317/622 , C12N2310/10
Abstract: Described herein are methods using CRISPR-Cas9 and DNA templates that can generate chimeric antigen receptors (CARs) on T cells to target the cell surface protein urokinase Plasminogen Activator Receptor (uPAR) on senescent cells. Also described are methods of preparing CAR T cells, their use to treat neurodegenerative disease, stroke, craniocerebral trauma and/or accident, or elderly individuals in need of treatment for aging.
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8.发明公开公开(公告)号：US20240076660A1
公开(公告)日：2024-03-07
申请号：US18348573
申请日：2023-07-07
Applicant: Wisconsin Alumni Research Foundation
IPC: C12N15/11 , A61K31/7088 , A61K35/545 , A61P3/00 , C12N9/22 , C12N15/86
CPC classification number: C12N15/11 , A61K31/7088 , A61K35/545 , A61P3/00 , C12N9/22 , C12N15/86 , C12N2310/20 , C12N2310/351 , C12N2800/80
Abstract: Described herein are methods of modifying a target gene in a patient or in a patient-derived cell, wherein the patient has an autosomal recessive disorder with compound heterozygous mutations, the methods including delivering a first modified guide RNA, a second modified guide RNA, a Cas9 polypeptide, a biotin-binding molecule, a first biotinylated donor polynucleotide, and a second biotinylated donor polynucleotide. The first modified guide RNA and the first biotinylated donor polynucleotide correct a first diseased allele, and the second modified guide RNA and the second biotinylated donor polynucleotide correct a second diseased allele.
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公开(公告)号：US20230272357A1
公开(公告)日：2023-08-31
申请号：US17980883
申请日：2022-11-04
Applicant: Wisconsin Alumni Research Foundation
Inventor： Krishanu Saha , Kaivalya Molugu
CPC classification number: C12N9/22 , C12N15/102 , A61P43/00 , C12N2310/20
Abstract: Described herein is an ex vivo method of site-specifically editing a target cell genome, the method including treating a population of unmodified target cells with a Class I and/or Class II histone deacetylase inhibitor to provide a population of chromatin decondensed unmodified target cells; and introducing into the population of chromatin decondensed unmodified target cells a Cas9 ribonucleoprotein, to provide a population of site-specifically genome-edited target cells; wherein the Cas9 ribonucleoprotein comprises a Cas9 protein and a guide RNA and cleaves DNA at a cleavage site in the target cell genome.
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公开(公告)号：US20230123017A1
公开(公告)日：2023-04-20
申请号：US17968027
申请日：2022-10-18
Applicant: WISCONSIN ALUMNI RESEARCH FOUNDATION
Inventor： Melissa C. Skala , Kaivalya Molugu , Krishanu Saha
Abstract: Systems and methods for identifying a current reprogramming status and for predicting a future reprogramming status for reprogramming intermediate cells (i.e., somatic cells undergoing reprogramming) are provided. Label-free autofluorescence measurements are combined with machine learning techniques to provide highly accurate identification of current reprogramming status and prediction of future reprogramming status. The identification of current reprogramming status utilizes metabolic endpoints from the autofluorescence data set. The prediction of future reprogramming status utilizes a pseudotime line constructed from autofluorescence data of reprogramming intermediate cells having a known reprogramming status.
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