Beam Therapeutics Inc.

Harvard College

The Broad Institute, Inc.

Other

Ipsen Biopharm Limited

Michael S Packer

Nicole Marie Gaudelli

Yi Yu

Bernd Zetsche

Ian Slaymaker

Seung-Joo Lee

David A Born

David R Liu

Ahmed Hussein Badran

Jason Michael Gehrke

Bryan Dickinson

J Keith Joung

Keith Foster

Matthew Beard

Gregory Newby

John Evans

Natalie Petrossian

Angelica Messana

Yvonne Aratyn

David Bryson

Francine Gregoire

Yanfang Fu

Genesis Lung

Travis R Blum

Shaunna Berkovitch

Provided herein are compositions and methods of using base editors comprising a polynucleotide programmable nucleotide binding domain and a nucleobase editing domain in conjunction with a guide polynucleotide. Also provided herein are base editor systems for editing nucleobases of target nucleotide sequences.

Methods of substituting pathogenic amino acids using programmable base editor systems

The disclosure provides amino acid sequence variants of Botulinum neurotoxin (BoNT) proteases that cleave (VAMP1, VAMP2, VAMP7, VAMP8, SNAP25, SNAP23, PTEN, etc.) and methods of evolving the same. In some embodiments, proteases described by the disclosure are useful for cleaving proteins found in a cell, that is in an intracellular environment. In some embodiments, proteases described by the disclosure are useful for treating diseases associated with increased or aberrant VAMP7, VAMP8, SNAP23 or PTEN expression or activity, for example, cancer and neurological disorders. Some aspects of this disclosure provide methods for generating BoNT protease variants by continuous directed evolution.

Evolution of BoNT peptidases

The present invention features compositions and methods for editing deleterious mutations associated with hemoglobinopathies, such as sickle cell disease (SCD). In particular embodiments, the invention provides methods for correcting mutations in a beta globin polynucleotide using modified adenosine base editors termed 'ABE8' having unprecedented levels (e.g., >60-70%) of efficiency.

Compositions and methods for treating hemoglobinopathies

The disclosure provides methods and compositions for treating blood diseases/disorders, such as sickle cell disease, hemochromatosis, hemophilia, and beta-thalassemia. For example the disclosure provides therapeutic guide RNAs that target the promotor of HBG1/2 to generate point mutations that increase expression of fetal hemoglobin. As another example, the disclosure provides therapeutic guide RNAs that target mutations in HBB, Factor VIII, and HFE to treat sickle cell disease, beta-thalassemia, hemophilia and hemochromatosis. The disclosure also provides fusion proteins comprising a Cas9 (e.g., a Cas9 nickase) domain and adenosine deaminases that deaminate adenosine in DNA. In some embodiments, the fusion proteins are in complex with nucleic acids, such as guide RNAs (gRNAs), which target the fusion proteins to a DNA sequence (e.g., an HBG1 or HBG2 protmoter sequence, or an HFE, GBB, or F8 gene sequence). Such complexes may be useful for increasing expression of fetal hemoglobin or correcting a poing mutation (e.g., C282Y) in HFE.

Uses of adenosine base editors

Some aspects of this disclosure provide methods for phage-assisted continuous evolution (PACE) of proteases. Some aspects of this invention provide methods for evaluating and selecting protease inhibitors based on the likelihood of the emergence of resistant proteases as determined by the protease PACE methods provided herein. Some aspects of this disclosure provide strategies, methods, and reagents for protease PACE, including fusion proteins for translating a desired protease activity into a selective advantage for phage particles encoding a protease exhibiting such an activity and improved mutagenesis-promoting expression constructs. Evolved proteases that recognize target cleavage sites which differ from their canonical cleavage site are also provided herein.

Evolution of proteases

The present invention features compositions and methods for editing deleterious mutations associated with hemoglobinopathies, such as sickle cell disease (SCD). In particular embodiments, the invention provides methods for correcting mutations in a beta globin polynucleotide using modified adenosine base editors termed 'ABE8' having unprecedented levels (e.g., &#x3e;60-70%) of efficiency.

The disclosure provides compositions comprising novel adenosine base editors (e.g., ABE8) that have increased efficiency and methods of using these adenosine deaminase variants for editing a target sequence.

Adenosine deaminase base editors and methods of using same to modify a nucleobase in a target sequence

A pedal for a percussion instrument, such as a bass drum, has a base plate and a foot plate pivotally mounted the base plate which is operably connected to an instrument striker. The pedal either has no heel plate or has a very short heel guide and is adapted so that when the tip of the player's foot is at an optimal position or 'sweet spot' on the foot plate, the player's heel extends to the playing surface.

Pedal for playing an instrument and pedal components

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Cambridge, MA, United States of America

Michael S Packer