A5015131912- Monoclonal and Polyclonal Antibodies Research
- Nanoparticle-Based Drug Delivery
- CRISPR and Genetic Engineering
- RNA Interference and Gene Delivery
- Advanced biosensing and bioanalysis techniques
- RNA and protein synthesis mechanisms
- Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Protein purification and stability
- Enzyme Structure and Function
- Bacterial Genetics and Biotechnology
- Radiopharmaceutical Chemistry and Applications
- DNA and Nucleic Acid Chemistry
- Biosimilars and Bioanalytical Methods
- Microbial Metabolic Engineering and Bioproduction
- Protein Structure and Dynamics
- Chemical Synthesis and Analysis
- Diet, Metabolism, and Disease
- Synthesis and Catalytic Reactions
- Liver Disease Diagnosis and Treatment
- Advanced Biosensing Techniques and Applications
- Advanced Drug Delivery Systems
- DNA Repair Mechanisms
- S100 Proteins and Annexins
- Glycosylation and Glycoproteins Research
- Trace Elements in Health
MedinCell (France)
2016-2022
Centre National de la Recherche Scientifique
1999-2012
Spanish National Cancer Research Centre
2010-2012
Inserm
2012
Hôpital Necker-Enfants Malades
2012
KU Leuven
2012
Université Paris Cité
2006-2012
CIC bioGUNE
2009-2010
Ikerbasque
2010
Délégation Paris 5
2008
Meganucleases, or homing endonucleases (HEs) are sequence-specific with large (>14 bp) cleavage sites that can be used to induce efficient homologous gene targeting in cultured cells and plants. These findings have opened novel perspectives for genome engineering a wide range of fields, including therapy. However, the number identified HEs does not match diversity genomic sequences, probability finding site chosen is extremely low. Therefore, design artificial specificities under intense...
Sequence-specific endonucleases recognizing long target sequences are emerging as powerful tools for genome engineering. These could be used to correct deleterious mutations or inactivate viruses, in a new approach molecular medicine. However, such applications highly demanding terms of safety. Mutations the human RAG1 gene cause severe combined immunodeficiency (SCID). Using I-CreI dimeric LAGLIDADG meganuclease scaffold, we describe here engineering series cleaving gene, including obligate...
A heterodimer of prenylated Rac1 and Rho GDP dissociation inhibitor was purified found to be competent in NADPH oxidase activation. Small angle neutron scattering experiments confirmed a 1:1 stoichiometry. The crystal structure the Rac1-RhoGDI complex determined at 2.7 resolution. In this which is bound GDP, switch I region conformation whereas II resembles that GTP-bound GTPase. Two types interaction between RhoGTPases RhoGDI were investigated. lipid-protein geranylgeranyl moiety resulted...
RmpM is a putative peptidoglycan binding protein from Neisseria meningitidis that has been shown to interact with integral outer membrane proteins such as porins and TonB-dependent transporters. Here we report the 1.9 A crystal structure of C-terminal domain RmpM. The 150-residue adopts betaalphabetaalphabetabeta fold, first identified in Bacillus subtilis chorismate mutase. homologous periplasmic, Escherichia coli OmpA; these domains are thought be responsible for non-covalent interactions...
The development of long-acting injectables (LAIs) for protein and peptide therapeutics has been a key challenge over the last 20 years. If these molecules offer advantages due to their high specificity selectivity, controlled release may confer several additional benefits in terms extended half-life, local delivery, patient compliance.This manuscript aims give an overview protein-based LAIs from industrial perspective, describing both approved promising technologies (with exceptions...
The ability to specifically engineer the genome of living cells at precise locations using rare-cutting designer endonucleases has broad implications for biotechnology and medicine, particularly functional genomics, transgenics gene therapy. However, potential impact chromosomal context epigenetics on endonuclease-mediated editing is poorly understood. To address this question, we conducted a comprehensive analysis efficacy 37 derived from quintessential I-CreI meganuclease that were...
The low molecular weight GTP binding protein Rac is essential to the activation of NADPH oxidase complex, involved in pathogen killing during phagocytosis. In resting cells, exists as a heterodimeric complex with Rho GDP dissociation inhibitor (Rho-GDI). Two types interactions exist between and Rho-GDI: protein−lipid interaction, implicating polyisoprene GTPase, well protein−protein interactions. Using two-hybrid system, we show that nonprenylated Rac1 interacts very weakly Rho-GDI, pointing...
Upon activation, the NADPH oxidase from neutrophils produces superoxide anions in response to microbial infection. This enzymatic complex is activated by association of its cytosolic factors p67phox , p47phox and small G protein Rac with a membrane-associated flavocytochrome b558. Here we report crystal structure active N-terminal fragment at 1.8 Å resolution, as well functional studies mutants. region (residues 1–213) consists mainly four TPR (tetratricopeptide repeat) motifs which C...
Homing endonucleases recognize long target DNA sequences generating an accurate double-strand break that promotes gene targeting through homologous recombination. We have modified the homodimeric I-CreI endonuclease protein engineering to a specific sequence within human RAG1 gene. Mutations in produce severe combined immunodeficiency (SCID), monogenic disease leading defective immune response individuals, leaving them vulnerable infectious diseases. The structures of two engineered...
Homing endonucleases have become valuable tools for genome engineering. Their sequence recognition repertoires can be expanded by modifying their specificities or creating chimeric proteins through domain swapping between two subdomains of different homing endonucleases. Here, we show that these approaches combined to create engineered meganucleases with new specificities. We demonstrate the modularity DmoCre meganuclease previously described, successfully assembling mutants locally altered...
Small therapeutic proteins represent a promising novel approach to treat cancer. Nevertheless, their clinical application is often adversely impacted by short plasma half-life. Controlled long-term delivery of small biologicals has become challenge because hydrophilic properties and in some cases limited stability. Here, an situ forming depot-injectable polymeric system was used deliver BiJ591, bispecific T-cell engager (BiTE) targeting both prostate-specific membrane antigen (PSMA) the CD3...
Abstract Heptosides are found in important bacterial glycolipids such as lipopolysaccharide (LPS), the biosynthesis of which is targeted for development novel antibacterial agents. This work describes synthesis a fluorinated analogue ADP‐ L ‐ glycero ‐β‐ D manno ‐heptopyranose, donor substrate heptosyl transferase WaaC, catalyzes incorporation this carbohydrate into LPS. Synthetically, key step preparation ADP‐2F‐heptose simultaneous and stereoselective installation both fluorine atom at C‐2...
Homing endonucleases, also known as meganucleases, are sequence-specific enzymes with large DNA recognition sites. These can be used to induce efficient homologous gene targeting in cells and plants, opening perspectives for genome engineering applications a wide series of fields, ranging from biotechnology therapy. Here, we report the crystal structures at 2.0 2.1 Å resolution I-DmoI meganuclease complex its substrate before after cleavage, providing snapshots catalytic process. Our study...
Homing endonucleases represent protein scaffolds that provide powerful tools for genome manipulation, as these enzymes possess a very low frequency of DNA cleavage in eukaryotic genomes due to their high specificity. The basis protein–DNA recognition must be understood generate tailored target the at sites interest. Protein–DNA interaction engineering homing has demonstrated potential approaches create new specific instruments genes inactivation or repair. interface studies have been focused...
Homing endonucleases (HE) have emerged as precise tools for achieving gene targeting events. Redesigned HEs with tailored specificities can be used to cleave new sequences, thereby considerably expanding the number of targetable genes and loci. With HEs, well other protein scaffolds, context dependence DNA/protein interaction patterns remains one major limitations rational engineering DNA binders. Previous studies shown strong crosstalk between different residues regions binding interface....
The NADPH oxidase of phagocytic cells is regulated by the cytosolic factors p47phox, p67phox, and p40phox as well Rac1−Rho-GDI heterodimer. regulation a consequence protein−protein interactions involving variety protein domains that are characterized in signal transduction. We have studied behavior solution using small angle neutron scattering gel filtration. two truncated forms namely, p47phox without its C-terminal end (residues 1−358) N-terminal 147−390), were found to be monomeric...
Ku70, a regulatory component of the DNA-dependent protein kinase, was identified by yeast two-hybrid screen B lymphocyte cDNA library as partner p40phox, O2--producing NADPH oxidase. Truncated constructs p40phox and Ku70 were used to map interacting sites. The 186 C-terminal amino acids (aa) found interact with two distinct regions central core region (aa 50-260) extremity 260-339). In complementary experiments, it observed that binds immobilized recombinant fusion from cell extract...
The generation of acylated impurities has represented an important hurdle in the development long acting injectables for therapeutic peptides using biocompatible polymers with a polyester moiety. We investigated here situ forming depot (ISFD) technology that uses polyethylene glycol - copolymers and solvent exchange mechanism to promote formation. This shown promise formulating small molecules as well proteins. In present work, well-known somatostatin analog octreotide acetate (OctAc) model...
Antisense-mediated exon skipping constitutes a promising new modality for treatment of Duchenne Muscular Dystrophy (DMD), which is caused by gene mutations that typically introduce translation stop codon in the dystrophin gene, thereby abolishing production functional protein. The removal can restore to produce shortened, but still partially Peptide nucleic acid (PNA) as potential antisense drug has previously been shown expression splice modulation mdx mouse model DMD. In this study, we...