CRISPR-Cas9 gene editing has emerged as a powerful therapeutic technology, but the lack of safe and efficient in vivo delivery systems, especially for tissue-specific vectors, limits its broad clinical applications. Delivery Cas9 ribonucleoprotein (RNP) owns competitive advantages over other options; however, large size RNPs exceeds loading capacity currently available vectors. Here, we report previously unidentified genome system, named exosome RNP , which were loaded into purified exosomes...

Transdermal delivery of genome-editing agents by microneedle promotes the glucocorticoid therapy inflammatory skin disorders.

The management of myocardial ischemia/reperfusion (I/R) damage in the context reperfusion treatment remains a significant hurdle field cardiovascular disorders. injured lesions exhibit distinctive features, including abnormal accumulation necrotic cells and subsequent inflammatory response, which further exacerbates impairment cardiac function. Here, we report genetically engineered hybrid nanovesicles (hNVs), contain cell-derived overexpressing high-affinity SIRPα variants (SαV-NVs),...

MOCAT is a highly configurable, modular pipeline for fast, standardized processing of single or paired-end sequencing data generated by the Illumina platform. The uses state-of-the-art programs to quality control, map, and assemble reads from metagenomic samples sequenced at depth several billion base pairs, predict protein-coding genes on assembled metagenomes. Mapping against reference databases allows read extraction removal, as well abundance calculations. Relevant statistics each step...

We synthesized a series of poly(disulfide)s by ring-opening polymerization and demonstrated that the copolymerization monomer 1 containing diethylenetriamine moieties 2 guanidyl ligands could generate an efficient delivery platform for different forms CRISPR-Cas9-based genome editors, including plasmid, mRNA, protein. The excellent performance designed stems from their delicate molecular structures to interact with genome-editing biomacromolecules, unique pathways mediate cellular uptake...

Regulation of CRISPR-Cas9 functions in vivo is conducive to developing precise therapeutic genome editing. Here, we report a prodrug nanosystem (termed NanoProCas9), which combines the targeted delivery and conditional activation for precision therapy inflammatory bowel disease. NanoProCas9 composed (i) cationic poly(β-amino ester) (PBAE) capable complexing plasmid DNA encoding destabilized Cas9 (dsCas9) nuclease, (ii) layer biomimetic cell membrane coated on PBAE/plasmid nanocomplexes...

Based on the high frequency of concurrent adenomatous polyposis coli (APC) and KRAS mutations their strong cooperative interaction in human colorectal cancer (CRC) promotion, we herein develop a CRISPR-Cas9-based genome-editing nanomedicine to target both APC for treatment CRC. To this end, hyaluronic acid (HA)-decorated phenylboronic dendrimer (HAPD) was designed targeted delivery Cas9 ribonucleoprotein (RNP), by which genetic harboring CRC cells can be synergistically disrupted. Systemic...

Developing protein drugs that can target intracellular sites remains a challenge due to their inadequate membrane permeability. Efficient carriers for cytosolic delivery are required protein-based drugs, cancer vaccines, and CRISPR-Cas9 gene therapies. Here, we report screening process identify highly efficient materials from library of dual-functionalized polymers bearing both boronate lipoic acid moieties. Both ligands were found be crucial binding, endosomal escape, release. Polymers with...

Abstract We synthesize supramolecular poly(disulfide) (CPS) containing covalently attached cucurbit[7]uril (CB[7]), which is exploited not only as a carrier to deliver plasmid DNA encoding destabilized Cas9 (dsCas9), but also host include trimethoprim (TMP) by CB[7] moieties through the complexation form TMP@CPS/dsCas9. Once transfected into tumor cells CPS, presence of polyamines can competitively trigger decomplexation TMP@CPS, thereby displacing and releasing TMP from stabilize dsCas9...

Bacteria lack an endoplasmic reticulum, a Golgi apparatus, and transport vesicles yet are capable of sorting delivering integral membrane proteins to particular sites within the cell with high precision. What is pathway by which reach their proper subcellular destination in bacteria? We have addressed this question using green fluorescent protein (GFP) fused polytopic (SpoIVFB) that involved process sporulation bacterium Bacillus subtilis. SpoIVFB-GFP localizes region sporulating known as...

Abstract Background Influenza A viruses (IAVs) are zoonotic, segmented negative-stranded RNA viruses. The rapid mutation of IAVs results in host immune response escape and antiviral drug vaccine resistance. RUNX1 is a transcription factor that not only plays essential roles hematopoiesis, but also functions as regulator inflammation. However, its role the innate immunity to IAV infection has been well studied. Methods To investigate effects on explore mechanisms uses during infection. We...

Intracellular delivery of therapeutic biomacromolecules is often challenged by the poor transmembrane and limited endosomal escape. Here, we establish a combinatorial library composed 150 molecular weight-defined gemini amphiphiles (GAs) to identify vehicles that facilitate robust cytosolic proteins in vitro vivo. These GAs display similar skeletal structures but differential amphiphilicity adjusting length alkyl tails, type ionizable cationic heads, hydrophobicity or hydrophilicity spacer....

Gene segments from other organisms, such as viruses, are detected foreign and targeted for silencing by RNAi pathways. A deep-sequencing map of the small RNA response to repeated transgenes introduced Caenorhabditis elegans revealed that specific siRNAs. Silencing gene depends on an antiviral involves changes in active silent chromatin modifications altered levels antisense Distinct Argonaute proteins target genes or protection against silencing. We used a transgene genome-wide screen...

Plant Trihelix transcription factors, specifically bind to GT elements and play important roles in plant physiology development. Wheat is a main cereal crop. Brachypodium distachyon close relative of wheat has been described as new model species for studying grass functional genomics. Presently, little known about B. genes. In 51 species, 2387 genes were identified, including 80 27 Consistent with the results previous studies, these classified into five subfamilies: GT-1, GT-2, SIP1, GTγ,...

Regulation of the cell-specific transcription factor sigma(F) in spore-forming bacterium Bacillus subtilis involves antisigma SpoIIAB. Contributing to activation is degradation SpoIIAB a manner that depends on protease ClpCP. Here we show three residues (LCN) located at extreme C terminus are both necessary and sufficient for this degradation. We also report use LCN extension as signal ClpCP unique

Gene expression late during the process of sporulation in Bacillus subtilis is governed by a multistep, signal transduction pathway involving transcription factor sigma(K), which derived regulated proteolysis from inactive proprotein pro-sigma(K). Processing pro-sigma(K) triggered signaling protein known as SpoIVB, serine protease that contains region with similarity to PDZ family protein-protein interaction domains. Here we report discovery second PDZ-containing called CtpB contributes...

Rational:The complex vascular architecture and diverse immune cells of the liver are critical for maintaining homeostasis.However, quantification network vasculature immunocytes at different scales from a single hepatic lobule to an intact lobe remains challenging.Methods: Here, we developed fast fluorescence-preserving transparency method, denoted liver-CUBIC, systematic integrated analysis microcirculation three-dimensional distribution dendritic (DCs)/macrophages in lobes.Results:...

Abstract We synthesize supramolecular poly(disulfide) (CPS) containing covalently attached cucurbit[7]uril (CB[7]), which is exploited not only as a carrier to deliver plasmid DNA encoding destabilized Cas9 (dsCas9), but also host include trimethoprim (TMP) by CB[7] moieties through the complexation form TMP@CPS/dsCas9. Once transfected into tumor cells CPS, presence of polyamines can competitively trigger decomplexation TMP@CPS, thereby displacing and releasing TMP from stabilize dsCas9...