A5033255466- Metal-Catalyzed Oxygenation Mechanisms
- Porphyrin Metabolism and Disorders
- CRISPR and Genetic Engineering
- Synthesis and Catalytic Reactions
- Microbial metabolism and enzyme function
- Catalytic C–H Functionalization Methods
- Redox biology and oxidative stress
- Chemical Synthesis and Analysis
- Enzyme Structure and Function
- Scientific Computing and Data Management
- GaN-based semiconductor devices and materials
- Virus-based gene therapy research
- CO2 Reduction Techniques and Catalysts
- Click Chemistry and Applications
- Insect symbiosis and bacterial influences
- Semiconductor Quantum Structures and Devices
- Cytomegalovirus and herpesvirus research
- RNA regulation and disease
- Metal complexes synthesis and properties
- Organic Light-Emitting Diodes Research
- Ubiquitin and proteasome pathways
- Hemoglobin structure and function
- Heme Oxygenase-1 and Carbon Monoxide
- Enzyme Catalysis and Immobilization
- Electron Spin Resonance Studies
Broad Institute
2022-2024
Harvard University
2021-2024
Howard Hughes Medical Institute
2023-2024
University of California, Berkeley
2019-2023
Center for Systems Biology
2021-2023
Abstract Cytosine base editors (CBEs) are larger and can suffer from higher off-target activity or lower on-target editing efficiency than current adenine (ABEs). To develop a CBE that retains the small size, low high of ABEs, we evolved highly active deoxyadenosine deaminase TadA-8e to perform cytidine deamination using phage-assisted continuous evolution. Evolved TadA deaminases contain mutations at DNA-binding residues alter enzyme selectivity strongly favor deoxycytidine over...
Prime editing enables a wide variety of precise genome edits in living cells. Here we use protein evolution and engineering to generate prime editors with reduced size improved efficiency. Using phage-assisted evolution, efficiencies compact reverse transcriptases by up 22-fold generated that are 516–810 base pairs smaller than the current-generation editor PEmax. We discovered different specialize types used this insight outperform PEmax PEmaxΔRNaseH, truncated dual-AAV delivery systems....
Abstract TadA-derived cytosine base editors (TadCBEs) enable programmable C•G-to-T•A editing while retaining the small size, high on-target activity, and low off-target activity of TadA deaminases. Existing TadCBEs, however, exhibit residual A•T-to-G•C at certain positions lower efficiencies some sequence contexts with non-SpCas9 targeting domains. To address these limitations, we use phage-assisted evolution to evolve CBE6s from a TadA-mediated dual adenine editor, discovering mutations N46...
Biocatalytic C-H activation has the potential to merge enzymatic and synthetic strategies for bond formation. FeII/αKG-dependent halogenases are particularly distinguished their ability both control selective as well direct group transfer of a bound anion along reaction axis separate from oxygen rebound, enabling development new transformations. In this context, we elucidate basis selectivity enzymes that perform halogenation yield 4-Cl-lysine (BesD), 5-Cl-lysine (HalB), 4-Cl-ornithine...
The enzyme BesC from the β-ethynyl-l-serine biosynthetic pathway in Streptomyces cattleya fragments 4-chloro-l-lysine (produced l-Lysine by BesD) to ammonia, formaldehyde, and 4-chloro-l-allylglycine can analogously fragment l-Lys itself. belongs emerging family of O2-activating non-heme-diiron enzymes with "heme-oxygenase-like" protein fold (HDOs). Here, we show that binding or an analogue triggers capture O2 protein's diiron(II) cofactor form a blue μ-peroxodiiron(III) intermediate...
An aliphatic halogenase requires four substrates: 2-oxoglutarate (2OG), halide (Cl
An aliphatic halogenase requires four substrates: 2-oxoglutarate (2OG), halide (Cl - or Br ), the halogenation target ("prime substrate"), and dioxygen. In well-studied cases, three non-gaseous substrates must bind to activate enzyme's Fe(II) cofactor for efficient capture of O 2 . Halide, 2OG, (lastly) all coordinate directly initiate its conversion a cis -halo-oxo-iron(IV) (haloferryl) complex, which abstracts hydrogen (H•) from non-coordinating prime substrate enable radicaloid...
ABSTRACT The enzyme BesC from the β - e thynyl-L- s erine biosynthetic pathway in Streptomyces cattleya fragments 4-chloro-L-lysine (produced L-Lysine by BesD) to ammonia, formaldehyde, and 4-chloro-L-allylglycine can analogously fragment L-Lys itself. belongs emerging family of O 2 -activating non-heme-diiron enzymes with “heme-oxygenase-like” protein fold (HDOs). Here we show that binding or an analog triggers capture protein’s diiron(II) cofactor form a blue µ-peroxodiiron(III)...
ABSTRACT Biocatalytic C–H activation has the potential to merge enzymatic and synthetic strategies for bond formation. Fe II /αKG-dependent halogenases are particularly distinguished their ability both control selective C-H as well direct group transfer of a bound anion along reaction axis separate from oxygen rebound, enabling development new transformations. In this context, we elucidate basis selectivity enzymes that perform halogenation yield 4-Cl-lysine (BesD), 5-Cl-lysine (HalB),...