A5003189849- Insect Resistance and Genetics
- CRISPR and Genetic Engineering
- Insect and Pesticide Research
- Neurobiology and Insect Physiology Research
- Insect symbiosis and bacterial influences
- RNA Interference and Gene Delivery
- RNA and protein synthesis mechanisms
- RNA Research and Splicing
- Studies on Chitinases and Chitosanases
- Invertebrate Immune Response Mechanisms
- Environmental Toxicology and Ecotoxicology
- Insect Pest Control Strategies
- Insect and Arachnid Ecology and Behavior
- Insect-Plant Interactions and Control
- Insect Utilization and Effects
- Lepidoptera: Biology and Taxonomy
- RNA regulation and disease
- Genetic diversity and population structure
- Orthoptera Research and Taxonomy
- Physiological and biochemical adaptations
- Bacterial Genetics and Biotechnology
- Species Distribution and Climate Change
- Pesticide Exposure and Toxicity
- Plant and animal studies
- RNA modifications and cancer
Shanxi University
2014-2023
Ministry of Education of the People's Republic of China
2023
University of California, Berkeley
2011-2022
Innovative Genomics Institute
2022
Shanxi University of Traditional Chinese Medicine
2011-2013
Howard Hughes Medical Institute
2009-2013
Kansas State University
2010
Institute of Molecular and Cell Biology
2005
Yale University
1997-2002
Medical College of Wisconsin
1994-1995
CRISPR-Cas12a (Cpf1) proteins are RNA-guided enzymes that bind and cut DNA as components of bacterial adaptive immune systems. Like CRISPR-Cas9, Cas12a has been harnessed for genome editing on the basis its ability to generate targeted, double-stranded breaks. Here we show binding unleashes indiscriminate single-stranded (ssDNA) cleavage activity by completely degrades ssDNA molecules. We find target-activated, nonspecific deoxyribonuclease (ssDNase) is also a property other type V...
Type II CRISPR immune systems in bacteria use a dual RNA-guided DNA endonuclease, Cas9, to cleave foreign at specific sites. We show here that Cas9 assembles with hybrid guide RNAs human cells and can induce the formation of double-strand breaks (DSBs) site complementary RNA sequence genomic DNA. This cleavage activity requires both binding RNA. Experiments using extracts from transfected expression and/or assembly into is limiting factor for Cas9-mediated cleavage. In addition, we find...
Type II CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems use an RNA-guided DNA endonuclease, Cas9, to generate double-strand breaks in invasive during adaptive bacterial immune response. Cas9 has been harnessed as a powerful tool for genome editing and gene regulation many eukaryotic organisms. We report 2.6 2.2 angstrom resolution crystal structures of two major enzyme subtypes, revealing the structural core shared by all family members. The...
CRISPR-Cas systems provide microbes with adaptive immunity to infectious nucleic acids and are widely employed as genome editing tools. These tools use RNA-guided Cas proteins whose large size (950 1400 amino acids) has been considered essential their specific DNA- or RNA-targeting activities. Here we present a set of from uncultivated archaea that contain Cas14, family exceptionally compact nucleases (400 700 acids). Despite small size, Cas14 capable targeted single-stranded DNA (ssDNA)...
Dicer is a central enzyme in microRNA (miRNA) processing. We identified Dicer-independent miRNA biogenesis pathway that uses Argonaute2 (Ago2) slicer catalytic activity. In contrast to other miRNAs, miR-451 levels were refractory dicer loss of function but reduced MZago2 (maternal-zygotic) mutants. found pre-miR-451 processing requires Ago2 activity vivo. mutants showed delayed erythropoiesis could be rescued by wild-type or miR-451-duplex not catalytically dead Ago2. Changing the secondary...
Targeted gene silencing by RNAi requires the RNA-induced complex (RISC), whose core component is protein Argonaute (Ago) bound to a microRNA (miRNA) or an siRNA. In humans, Ago2 loaded with miRNAs action of specialized assembly called RISC-loading (RLC), comprising proteins Ago2, Dicer, and TRBP. Here we show that human RLC assembles spontaneously in vitro from purified components. No cofactors chaperones are required for form. The reconstituted RLC, containing one copy each protein, has...
CRISPR-Cas9 is a powerful technology that has enabled genome editing in wide range of species. However, the currently developed Cas9 homologs all originate from mesophilic bacteria, making them susceptible to degradation and unsuitable for applications requiring cleavage at elevated temperatures. Here, we show protein thermophilic bacterium Geobacillus stearothermophilus (GeoCas9) catalyzes RNA-guided DNA GeoCas9 active temperatures up 70 °C, compared 45 °C Streptococcus pyogenes (SpyCas9),...
Significance Efficient nonviral delivery of macromolecules including mRNA, DNA plasmids, Cas9 ribonucleoproteins, and functional protein into both adherent cells suspension with high cell viability is crucial for cellular manipulation, imaging, medical applications. However, the conventional methods are limited to a certain range molecules types often reduce viability. Here, we demonstrate effective by nanopore electroporation (NanoEP) using water-filter nanoporous membrane. As compared that...
Intracellular delivery of mRNA, DNA, and other large macromolecules into cells plays an essential role in array biological research clinical therapies. However, current methods yield a wide variation the amount material delivered, as well limitations on cell types cargoes possible. Here, we demonstrate quantitatively controlled range primary lines with tight dosage distribution using nanostraw-electroporation system (NES). In NES, are cultured onto track-etched membranes protruding...
Abstract CRISPR-Cas12a is an RNA-guided, programmable genome editing enzyme found within bacterial adaptive immune pathways. Unlike CRISPR-Cas9, Cas12a uses only a single catalytic site to both cleave target double-stranded DNA (dsDNA) (cis-activity) and indiscriminately degrade single-stranded (ssDNA) (trans-activity). To investigate how the relative potency of cis- versus trans-DNase activity affects Cas12a-mediated editing, we first used structure-guided engineering generate variants...
Recent studies have shown that trehalose plays a protective role in yeast variety of stresses, including heat, freezing and thawing, dehydration, hyperosmotic shock, oxidant injury. Because (a) heat shock anoxia share mechanisms allow organisms to survive, (b) Drosophila melanogaster is tolerant anoxia, (c) present flies metabolically active, we asked whether can protect against anoxic stress. Here report on new resistance Drosophila. We first cloned the gene trehalose-6-phosphate synthase...
Chitinase is an important enzyme responsible for chitin metabolism in a wide range of organisms including bacteria, yeasts and other fungi, nematodes arthropods. However, current knowledge on chitinolytic enzymes, especially their structures, functions regulation very limited. In this study we have identified 20 chitinase chitinase-like genes the African malaria mosquito, Anopheles gambiae, through genome-wide searching transcript profiling. We assigned these into eight different groupings...