公开(公告)号：US20210230568A1

公开(公告)日：2021-07-29

申请号：US17051632

申请日：2019-05-03

Applicant: UNIVERSITY OF MASSACHUSETTS

Inventor： Scot A. Wolfe ,  Charles P. Emerson, JR. ,  Sukanya Iyer ,  Sneha Suresh ,  Christian Mueller ,  Jennifer Chen ,  Dongsheng Guo ,  Oliver King

IPC: C12N9/22 ,  A61K35/76 ,  A61K38/00

Abstract: The present invention is directed to the filed of gene therapy. In particular, compositions and methods are disclosed that repair gene microduplication mutations by reversion to a wild type sequence. For example, the creation of a double stranded break by a programmable nuclease protein within a microduplication induces the microhomology mediated end joining DNA repair pathway that in the process of DNA repair removes the microduplication mutation and restores the wild type sequence.

公开(公告)号：US20190276810A1

公开(公告)日：2019-09-12

申请号：US16226202

申请日：2018-12-19

Applicant: University of Massachusetts

Inventor： Scot A. Wolfe ,  Mehmet Fatih Bolukbasi ,  Ankit Gupta ,  Erik J. Sontheimer ,  Nadia Amrani

IPC: C12N9/22

Abstract: The present invention provides a Cas9 platform to facilitate single-site nuclease gene editing precision within a human genome. For example, a Cas9 nuclease/DNA-targeting unit (Cas9-DTU) fusion protein precisely delivers a Cas9/sgRNA complex to a specific target site within the genome for subsequent sgRNA-dependent cleavage of an adjacent target sequence. Alternatively, attenuating Cas9 binding using mutations to the a protospacer adjacent motif (PAM) recognition domain makes Cas9 target site recognition dependent on the associated DTU, all while retaining Cas9's sgRNA-mediated DNA cleavage fidelity. Cas9-DTU fusion proteins have improved target site binding precision, greater nuclease activity, and a broader sequence targeting range than standard Cas9 systems. Existing Cas9 or sgRNA variants (e.g., truncated sgRNAs (tru-gRNAs), nickases and FokI fusions) are compatible with these improvements to further reduce off-target cleavage. A robust, broadly applicable strategy is disclosed to impart Cas9 genome-editing systems with the single-genomic-site accuracy needed for safe, effective clinical application.

公开(公告)号：US20240287486A1

公开(公告)日：2024-08-29

申请号：US17921027

申请日：2021-04-21

Applicant: UNIVERSITY OF MASSACHUSETTS

Inventor： Scot A. Wolfe ,  Kevin Luk ,  PengPeng Liu

IPC: C12N9/22 ,  A61K48/00 ,  C12N15/90

CPC classification number: C12N9/22 ,  A61K48/005 ,  C12N15/907 ,  C07K2319/09 ,  C12N2310/20 ,  C12N2800/80

Abstract: The present invention is related to the field of gene editing. In particular, the present invention is related to the mutation and/or deletion of genetic abnormalities that result in genetic diseases. For example, an improved CRISPR-Cas fusion protein is disclosed where the Cas protein is a Cas12a protein. The Cas12a protein is fused to a variety of nuclear localization signal (NLS) sequences (e.g., c-myc NLS) that are demonstrated to have unexpected and superior gene editing activity when compared to conventional NLS sequences (e.g., SV40 NLS).

公开(公告)号：US20230279405A1

公开(公告)日：2023-09-07

申请号：US18017479

申请日：2021-07-23

Applicant: University of Massachusetts

Inventor： Miguel Sena Esteves ,  Scot A. Wolfe

IPC: C12N15/62 ,  C12N15/86 ,  C12N15/10

CPC classification number: C12N15/62 ,  C12N15/86 ,  C12N15/1034 ,  C07K2319/01 ,  C12N2750/14143 ,  C12N2830/008

Abstract: In some aspects, the disclosure relates to recombinant adeno-associated viruses (rAAVs) comprising a nucleic acid encoding a fusion protein comprising a DNA-binding domain and a transcriptional regulator domain and methods of using the same. In some embodiments, expression of the fusion protein results in modified expression of a target gene in a cell.

公开(公告)号：US20220290113A1

公开(公告)日：2022-09-15

申请号：US17285440

申请日：2019-10-15

Applicant: UNIVERSITY OF MASSACHUSETTS

Inventor： Erik J. Sontheimer ,  Xin Gao ,  Aamir Mir ,  Alireza Edraki ,  Scot A. Wolfe ,  Pengpeng Liu

IPC: C12N9/22 ,  C12N9/78 ,  C12N15/11 ,  C12N15/62 ,  C12N15/86 ,  A61P3/00

Abstract: The present invention is related to the field of gene editing. In particular, the gene editing is directed toward single nucleotide base editing. For example, such single nucleotide base editing results in a conversion of a OG base pair to a T*A base pair. The high accuracy and precision of the presently disclosed single nucleotide base gene editor is accomplished by an NmeCas9 nuclease that is fused to a nucleotide deaminase protein. The compact nature of the NmeCas9 coupled with a larger number of compatible protospacer adjacent motifs provide the Cas9 fusion constructs contemplated herein to have a gene editing window that can edit sites that are not targetable by other conventional SpyCas9 base editor platforms.

公开(公告)号：US20240141359A1

公开(公告)日：2024-05-02

申请号：US18278338

申请日：2022-02-23

Applicant: UNIVERSITY OF MASSACHUSETTS

Inventor： Charles P. Emerson, JR. ,  Scot A. Wolfe

IPC: C12N15/113 ,  C12N9/22 ,  C12N15/86

CPC classification number: C12N15/1138 ,  C12N9/22 ,  C12N15/86 ,  C12N2310/20 ,  C12N2750/14143

Abstract: The present invention is related to the field of genetic engineering. In particular, the repair, reversion and/or conversion of genetic mutations that are linked to a muscular dystrophy disease. Specifically contemplated are gene editor nuclease proteins or base editor proteins that are targeted to the muscular dystrophy genetic mutations or pathogenic variants. Such gene editor nuclease proteins include, but are not limited to Cas12a nuclease proteins and adenine base editor proteins. Repair, reversion and/or disruption of the genetic mutation or pathogenic variant reduces at least one symptom of a muscular dystrophy disease.

公开(公告)号：US20230049455A1

公开(公告)日：2023-02-16

申请号：US17796184

申请日：2021-01-29

Applicant: UNIVERSITY OF MASSACHUSETTS

Inventor： Scot A. Wolfe ,  Pengpeng Liu ,  Kevin Luk

IPC: C12N9/22 ,  C12N15/11 ,  C12N15/62 ,  C12N9/78 ,  C12N15/90 ,  C07K14/195

Abstract: RNA-guided programmable cytosine and adenine base editors are a powerful class of genome editing tool for the introduction of localized base transitions without generating a double-stranded DNA break. Base editors (BE) have an optimal window of activity relative to the PAM recognized by the Cas9 enzyme and these constructs are strand selective. Here we demonstrate that fusion of a programmable DNA-binding domain (pDBD) or another Cas9 orthologue to spCas9-BE, we can produce an RNA-programmable Cas9-BE-pDBD chimera or Cas9-BE-Cas9 chimeras with dramatically improved activities and increased targeting range. Cas9-pDBD or Cas9-Cas9 fusion base editors display an expanded targeting repertoire and achieve highly specific genome editing, which can be tailored to achieve extremely precise genome editing at nearly any genomic locus.

公开(公告)号：US20220185862A1

公开(公告)日：2022-06-16

申请号：US17433269

申请日：2020-02-24

Applicant: University of Massachusetts

Inventor： Miguel Sena Esteves ,  Scot A. Wolfe

IPC: C07K14/705 ,  C12N9/22 ,  C12N15/11 ,  C12N15/86

Abstract: In some aspects, the disclosure relates to recombinant adeno-associated viruses (rAAVs) comprising a nucleic acid encoding a fusion protein comprising a DNA-binding domain and a transcriptional regulator domain and methods of using the same. In some embodiments, expression of the fusion protein results in modified expression of a target gene in a cell.