A5044251104- Porphyrin Metabolism and Disorders
- CRISPR and Genetic Engineering
- Metal-Catalyzed Oxygenation Mechanisms
- Folate and B Vitamins Research
- RNA regulation and disease
- Bacteriophages and microbial interactions
- Hemoglobinopathies and Related Disorders
- Enzyme Structure and Function
- Yersinia bacterium, plague, ectoparasites research
- Microbial bioremediation and biosurfactants
- Cytomegalovirus and herpesvirus research
- RNA Interference and Gene Delivery
- Biochemical and Molecular Research
- Metabolism and Genetic Disorders
- Enzyme Catalysis and Immobilization
- Microbial Natural Products and Biosynthesis
- Bacillus and Francisella bacterial research
- Advanced Breast Cancer Therapies
- Hepatitis B Virus Studies
- Amino Acid Enzymes and Metabolism
- Pharmacogenetics and Drug Metabolism
- Metal Extraction and Bioleaching
- Iron Metabolism and Disorders
- Catalysis and Hydrodesulfurization Studies
- Organometallic Compounds Synthesis and Characterization
Massachusetts Institute of Technology
2015-2023
Harvard University
2015-2023
Beam Therapeutics (United States)
2020-2021
Amgen (United States)
2019
Kalpavriksh
2015
Abstract Cytosine base editors (CBEs) enable efficient, programmable reversion of T•A to C•G point mutations in the human genome. Recently, cytosine with rAPOBEC1 were reported induce unguided deamination genomic DNA and cellular RNA. Here we report eight next-generation CBEs (BE4 either RrA3F [wt, F130L], AmAPOBEC1, SsAPOBEC3B R54Q], or PpAPOBEC1 H122A, R33A]) that display comparable on-target editing frequencies, whilst eliciting a 12- 69-fold reduction C-to-U edits transcriptome, up...
Abstract Only a small fraction of early drug programs progress to the market, due safety and efficacy failures, despite extensive efforts predict safety. Characterizing effect natural variation in genes encoding targets should present powerful approach side effects arising from drugging particular proteins. In this retrospective analysis, we report correlation between organ systems affected by genetic which are observed. Across 1819 drugs 21 phenotype categories analyzed, more likely occur...
Base editors are fusions of a deaminase and CRISPR-Cas ribonucleoprotein that allow programmable installment transition mutations without double-strand DNA break intermediates. The breadth potential base editing targets is frequently limited by the requirement suitably positioned Cas9 protospacer adjacent motif. To address this, we used structures TadA to design set inlaid (IBEs), in which domains internal Cas9. Several these IBEs exhibit shifted windows greater efficiency, enabling outside...
Methylphosphonate synthase (MPnS) produces methylphosphonate, a metabolic precursor to methane in the upper ocean. Here, we determine 2.35-angstrom resolution structure of MPnS and discover that it has an unusual 2-histidine-1-glutamine iron-coordinating triad. We further solve related enzyme, hydroxyethylphosphonate dioxygenase from Streptomyces albus (SaHEPD), find displays same motif. SaHEPD can be converted into by mutation glutamine-adjacent residues, identifying molecular requirements...
Abstract The electron‐rich isonitrile is an important functionality in bioactive natural products, but its biosynthesis has been restricted to the IsnA family of synthases. We herein provide first structural and biochemical evidence alternative mechanism for formation. ScoE, a putative non‐heme iron(II)‐dependent enzyme from Streptomyces coeruleorubidus , was shown catalyze conversion ( R )‐3‐((carboxymethyl)amino)butanoic acid )‐3‐isocyanobutanoic through oxidative decarboxylation...
Acyl-CoA mutases are a growing class of adenosylcobalamin-dependent radical enzymes that perform challenging carbon skeleton rearrangements in primary and secondary metabolism. Members this must precisely control substrate positioning to prevent oxidative interception intermediates during catalysis. Our understanding specificity catalysis acyl-CoA mutases, however, is incomplete. Here, we present crystal structures IcmF, natural fusion protein variant isobutyryl-CoA mutase, complex with the...
G-protein metallochaperone MeaB in bacteria [methylmalonic aciduria type A (MMAA) humans] is responsible for facilitating the delivery of adenosylcobalamin (AdoCbl) to methylmalonyl-CoA mutase (MCM), only AdoCbl-dependent enzyme humans. Genetic defects switch III region MMAA lead genetic disorder methylmalonic which body unable process certain lipids. Here, we present a crystal structure Methylobacterium extorquens bound nonhydrolyzable guanosine triphosphate (GTP) analog...
The foundational adenine base editors ( e.g. ABE7.10) enable programmable C•G to T•A point mutations but editing efficiencies can be low at challenging loci in primary human cells. Here we further evolve ABE7.10 using a library of adenosine deaminase variants create ABE8s. At NGG PAM sites, ABE8s result ∼1.5x higher protospacer positions A5-A7 and ∼3.2x A3-A4 A8-A10 compared with ABE7.10. Non-NGG have ∼4.2-fold overall on-target efficiency than In CD34+ cells, ABE8 recreate natural allele...
Abstract The electron‐rich isonitrile is an important functionality in bioactive natural products, but its biosynthesis has been restricted to the IsnA family of synthases. We herein provide first structural and biochemical evidence alternative mechanism for formation. ScoE, a putative non‐heme iron(II)‐dependent enzyme from Streptomyces coeruleorubidus , was shown catalyze conversion ( R )‐3‐((carboxymethyl)amino)butanoic acid )‐3‐isocyanobutanoic through oxidative decarboxylation...
G-protein metallochaperones are essential for the proper maturation of numerous metalloenzymes. The chaperone MMAA in humans (MeaB bacteria) uses GTP hydrolysis to facilitate delivery adenosylcobalamin (AdoCbl) AdoCbl-dependent methylmalonyl-CoA mutase, an metabolic enzyme. This also facilitates removal damaged cobalamin (Cbl) repair. Although most chaperones standalone proteins, isobutyryl-CoA mutase fused (IcmF) has a domain covalently attached its target mutase. We previously showed that...
Abstract/introductory paragraph Cytosine base editors (CBEs) are molecular machines which enable efficient and programmable reversion of T•A to C•G point mutations in the human genome without induction DNA double strand breaks 1, 2 . Recently, foundational cytosine editor (CBE) ‘BE3’, containing rAPOBEC1, was reported induce unguided, genomic 3, 4 cellular RNA 5 deamination when expressed living cells. To mitigate spurious off-target events, we developed a sensitive, high-throughput assay...
Abstract The electron-rich isonitrile is an important functionality in bioactive natural products, but its biosynthesis has been restricted to the IsnA family of synthases. We here provide first structural and biochemical evidence alternative mechanism for formation. ScoE, a putative non-heme iron(II)-dependent enzyme from Streptomyces coeruleorubidus , was shown catalyze conversion ( R )-3-((carboxymethyl)amino)butanoic acid )-3-isocyanobutanoic through oxidative decarboxylation mechanism....