A5041880278- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- Advanced biosensing and bioanalysis techniques
- Gastric Cancer Management and Outcomes
- Metallurgy and Material Forming
- Cancer-related gene regulation
- RNA regulation and disease
- Aluminum Alloy Microstructure Properties
- Metal Forming Simulation Techniques
- Esophageal and GI Pathology
- RNA modifications and cancer
- Pancreatic and Hepatic Oncology Research
- Cholangiocarcinoma and Gallbladder Cancer Studies
- Microstructure and mechanical properties
- Biochemical and Molecular Research
- Gallbladder and Bile Duct Disorders
- Eosinophilic Esophagitis
- Cancer Diagnosis and Treatment
- Gastrointestinal Tumor Research and Treatment
- Liver Disease Diagnosis and Treatment
- Silkworms and Sericulture Research
- Colorectal Cancer Treatments and Studies
- Systemic Sclerosis and Related Diseases
- Livestock and Poultry Management
- Retinopathy of Prematurity Studies
The University of Tokyo
2015-2024
Broad Institute
2022-2024
Howard Hughes Medical Institute
2023-2024
McGovern Institute for Brain Research
2023-2024
Massachusetts Institute of Technology
2023-2024
Hokkaido University
2018
Takatsuki General Hospital
2014-2017
Komaki City Hospital
2010
Tokyo University of Agriculture and Technology
2006
Kyoto first Red Cross hospital
1990-1998
The RNA-guided endonuclease Cas9 cleaves its target DNA and is a powerful genome-editing tool. However, the widely used Streptococcus pyogenes enzyme (SpCas9) requires an NGG protospacer adjacent motif (PAM) for recognition, thereby restricting targetable genomic loci. Here, we report rationally engineered SpCas9 variant (SpCas9-NG) that can recognize relaxed NG PAMs. crystal structure revealed loss of base-specific interaction with third nucleobase compensated by newly introduced...
A cap-specific m 6 writer N ,2′- O -dimethyladenosine (m Am) is present at the transcription start nucleotide of capped mRNAs in vertebrates. Akichika et al. quantified abundance this modification transcriptome and identified protein, adenosine methyltransferase (CAPAM), needed for modification. CAPAM contains a unique structure that recognizes -methyladenosine A) as substrate. The protein interacts with RNA polymerase II, suggesting occurs cotranscriptionally. Am promotes translation...
The CRISPR-associated endonuclease Cas9 binds to a guide RNA and cleaves double-stranded DNA with sequence complementary the guide. Cas9-RNA system has been harnessed for numerous applications, such as genome editing. Here we use high-speed atomic force microscopy (HS-AFM) visualize real-space real-time dynamics of CRISPR-Cas9 in action. HS-AFM movies indicate that, whereas apo-Cas9 adopts unexpected flexible conformations, forms stable bilobed structure interrogates target sites on by...
ENPP1 (Ecto-nucleotide pyrophosphatase phosphodiesterase 1), a type II transmembrane glycoprotein, hydrolyzes ATP to produce AMP and diphosphate, thereby inhibiting bone mineralization. A recent study showed that also preferentially 2'3'-cGAMP (cyclic GMP-AMP) but not its linkage isomer 3'3'-cGAMP, negatively regulates the cGAS-STING pathway in innate immune system. Here, we present high-resolution crystal structures of complex with 3'3'-cGAMP reaction intermediate pA(3',5')pG. The revealed...
TnpB is a member of the Obligate Mobile Element Guided Activity (OMEGA) RNA-guided nuclease family, harbored in transposons, and likely functions to maintain transposon genomes. Previously, it was shown that cleaves double- single-stranded DNA substrates an manner, but biogenesis ribonucleoprotein (RNP) complex unknown. Using
Abstract The clinical success of CRISPR therapies hinges on the safety and efficacy Cas proteins. Cas9 from Francisella novicida (FnCas9) is highly precise, with a negligible affinity for mismatched substrates, but its low cellular targeting efficiency limits therapeutic use. Here, we rationally engineer protein to develop enhanced FnCas9 (enFnCas9) variants broaden their accessibility across human genomic sites by ~3.5-fold. enFnCas9 proteins single mismatch specificity expanded target...
Abstract PIWI-clade Argonaute proteins associate with PIWI-interacting RNAs (piRNAs), and silence transposons in animal gonads. Here, we report the crystal structure of Drosophila Piwi complex endogenous piRNAs, at 2.9 Å resolution. A structural comparison other Argonautes highlights PIWI-specific features, such as overall domain arrangement metal-dependent piRNA recognition. Our biochemical data reveal that, unlike including silkworm Siwi, has a non-canonical DVDK tetrad lacks RNA-guided...
Abstract The RNA-guided DNA endonuclease Cas9 cleaves double-stranded targets bearing a protospacer adjacent motif (PAM) and complementarity to an RNA guide. Unlike other orthologs, Corynebacterium diphtheriae (CdCas9) recognizes the promiscuous NNRHHHY PAM. However, CdCas9-mediated PAM recognition mechanism remains unknown. Here, we report crystal structure of CdCas9 in complex with guide its target at 2.9 Å resolution. reveals that via combination van der Waals interactions base-specific...
RNA-guided systems, such as CRISPR-Cas, combine programmable substrate recognition with enzymatic function, a combination that has been used advantageously to develop powerful molecular technologies
Canonical prokaryotic type I CRISPR-Cas adaptive immune systems contain a multicomponent effector complex called Cascade, which degrades large stretches of DNA via Cas3 helicase-nuclease activity. Recently, highly precise subtype I-F1 system (HNH-Cascade) was found that lacks Cas3, the absence is compensated for by insertion an HNH endonuclease domain in Cas8 Cascade component. Here, we describe cryo-EM structure Selenomonas sp. HNH-Cascade (SsCascade) with target and characterize its...
Increasing incidence of various cancers has been reported in diabetic patients. O ‐linked N ‐acetylglucosamine ( ‐Glc NA c) modification proteins at serine/threonine residues cylation) is an essential post‐translational that upregulated patients and implicated tumor growth. However, the mechanisms by which cylation promotes growth remain unclear. Given AMP ‐activated kinase AMPK ) thought to play important roles suppressing growth, we evaluated involvement on LoVo cells, a human colon cancer...
We aimed to elucidate the risk factors and preventive associated with chronic low-dose aspirin (L-ASA)-induced gastroduodenal mucosal injury in Japanese patients arteriosclerotic disease.This retrospective observational study included 400 L-ASA users who underwent upper gastrointestinal endoscopy. investigated patients' clinical characteristics, including age, peptic ulcer history, concomitant drugs [i.e. gastric agents, antiplatelet drugs, anticoagulants, non-steroidal anti-inflammatory...
Forming Limit Curves (FLCs) of 5000 series aluminum alloys with different Mg contents, 2.5 and 5.5 mass%, are experimentally determined in detail using the Marciniak test. The forming limit strains Al–Mg 5.5% alloy larger than those 2.5% from uniaxial tension to near plane-strain regions, whereas order formability is reversed biaxial stretching region. To evaluate work-hardening behavior test materials under determine appropriate anisotropic yield functions, tensile tests carried out...
Two types of a shape memory alloy (SMA) catheter system with mechanism for tip control and irrigation were evaluated in performing peroral pancreatoscopy (PPS) using an ultrathin-caliber endoscope. The selective cannulation SMA catheters into the papilla was made possible 18 19 patients pancreatic diseases. Endoscopic sphincterotomy necessary six seven two-directional (2.6 mm) but only one 12 examined one-directional type (2.2 mm). In 17 patients, smoothly advanced to desired positions main...
<b>Background/Aims: </b>There have been few reliable published studies permitting assessment of therapeutic regimens for <i>Helicobacter pylori</i> infection in gastric ulcer (GU) patients. The aim the present study was to evaluate efficacy omeprazole-based dual and triple therapy regimens, both including clarithromycin, cure <i>H. active GU duodenal (DU) conducted Japan, a country which is more prevalent than DU. <b>Methods: </b>Two hundred...
The CRISPR-associated endonuclease Cas9 can be targeted to specific genomic loci by single guide RNAs (sgRNAs).We first solved the crystal structure of Streptococcus pyrogenes (SpCas9) in complex with sgRNA and its target DNA at 2.5 A resolution.The revealed a bilobed architecture consisting recognition nuclease lobes (Rec Nuc lobes, respectively), accommodating sgRNA:DNA heteroduplex positively-charged groove their interface.While Rec lobe is essential for binding DNA, contains HNH RuvC...
<title>Abstract</title> The clinical success of CRISPR therapies is dependent on the safety and efficacy Cas proteins. Cas9 from <italic>Francisella novicida</italic> (FnCas9) has negligible affinity for mismatched substrates enabling it to discriminate off-targets in DNA with very high precision even at level binding. However, its cellular targeting efficiency low, limiting use therapeutic applications. Here, we rationally engineer protein develop enhanced FnCas9 (enFnCas9) variants expand...