Pathogen Recognition and Innate Immunity

Bacteria and archaea have evolved adaptive immune defenses, termed clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems, that use short RNA to direct degradation of foreign nucleic acids. Here, we engineer the type II bacterial CRISPR system to function with custom guide RNA (gRNA) in human cells. For the endogenous AAVS1 locus, we obtained targeting rates of 10 to 25% in 293T cells, 13 to 8% in K562 cells, and 2 to 4% in induced pluripotent stem cells. We show that this process relies on CRISPR components; is sequence-specific; and, upon simultaneous introduction of multiple gRNAs, can effect multiplex editing of target loci. We also compute a genome-wide resource of ~190 K unique gRNAs targeting ~40.5% of human exons. Our results establish an RNA-guided editing tool for facile, robust, and multiplexable human genome engineering.

/pdf/rna-guided-human-genome-engineering-via-cas9-4eefu2kibr.pdf

RNA-Guided Human Genome Engineering via Cas9

The discovery of Toll-like receptors (TLRs) as components that recognize conserved structures in pathogens has greatly advanced understanding of how the body senses pathogen invasion, triggers innate immune responses and primes antigen-specific adaptive immunity. Although TLRs are critical for host defense, it has become apparent that loss of negative regulation of TLR signaling, as well as recognition of self molecules by TLRs, are strongly associated with the pathogenesis of inflammatory and autoimmune diseases. Furthermore, it is now clear that the interaction between TLRs and recently identified cytosolic innate immune sensors is crucial for mounting effective immune responses. Here we describe the recent advances that have been made by research into the role of TLR biology in host defense and disease.

/pdf/the-role-of-pattern-recognition-receptors-in-innate-immunity-39y77tlxza.pdf

The role of pattern-recognition receptors in innate immunity: update on Toll-like receptors

Pattern Recognition Receptors and Inflammation

The transdifferentiation of epithelial cells into motile mesenchymal cells, a process known as epithelial-mesenchymal transition (EMT), is integral in development, wound healing and stem cell behaviour, and contributes pathologically to fibrosis and cancer progression. This switch in cell differentiation and behaviour is mediated by key transcription factors, including SNAIL, zinc-finger E-box-binding (ZEB) and basic helix-loop-helix transcription factors, the functions of which are finely regulated at the transcriptional, translational and post-translational levels. The reprogramming of gene expression during EMT, as well as non-transcriptional changes, are initiated and controlled by signalling pathways that respond to extracellular cues. Among these, transforming growth factor-β (TGFβ) family signalling has a predominant role; however, the convergence of signalling pathways is essential for EMT.

Molecular mechanisms of epithelial–mesenchymal transition

Surface modulation of extracellular vesicles with cell-penetrating peptide-conjugated lipids for improvement of intracellular delivery to endothelial cells

Development of siRNA libraries by in vitro dicing and optimized efficient expression vectors for siRNAs in mammalian cells

Design of Intracellularly Active Ribozymes and siRNAs

p52-ZER6: a determinant of tumor cell sensitivity to MDM2-p53 binding inhibitors

L'invention concerne un ARN interference bicatenaire, qui comprend: un ARN antisens complementaire d'une sequence situee dans la region avoisinante d'un gene cible ou d'une sequence situee dans n'importe quelle region d'un ARNm du gene cible; et un ARN sens homologue de ladite sequence pouvant s'hybrider avec ledit ARN antisens dans une cellule. L'ARN antisens et l'ARN sens forment des doubles brins et le fragment bicatenaire presente une longueur constituee d'au moins 50 bases. L'utilisation de cet ARN interference bicatenaire permet d'induire de maniere tres efficace un ARNi de toxicite reduite, sans necessite de chercher a determiner la partie a utiliser comme sequence complementaire de l'ARN antisens. En introduisant une substitution de bases ou renflement dans un tel ARN interference bicatenaire long, l'ARNi peut etre ameliore encore et la reponse a l'interferon reduite.

Makoto Miyagishi

Papers

Surface modulation of extracellular vesicles with cell-penetrating peptide-conjugated lipids for improvement of intracellular delivery to endothelial cells

Development of siRNA libraries by in vitro dicing and optimized efficient expression vectors for siRNAs in mammalian cells

Design of Intracellularly Active Ribozymes and siRNAs

p52-ZER6: a determinant of tumor cell sensitivity to MDM2-p53 binding inhibitors

Arn bicatenaire long interferant avec une reponse de l'interferon reduite