A5072124571- CRISPR and Genetic Engineering
- Pluripotent Stem Cells Research
- RNA regulation and disease
- RNA and protein synthesis mechanisms
- Renal and related cancers
- RNA Interference and Gene Delivery
- Virus-based gene therapy research
- Reproductive Biology and Fertility
- Forensic and Genetic Research
- Retinal Development and Disorders
- Molecular Biology Techniques and Applications
- Pain Mechanisms and Treatments
- Cancer, Hypoxia, and Metabolism
- Cell death mechanisms and regulation
- Forensic Entomology and Diptera Studies
- Genetics, Aging, and Longevity in Model Organisms
- Genetics and Neurodevelopmental Disorders
- interferon and immune responses
- Anesthesia and Pain Management
- Paleopathology and ancient diseases
- Forensic Anthropology and Bioarchaeology Studies
- NF-κB Signaling Pathways
- Neuroscience of respiration and sleep
- Ubiquitin and proteasome pathways
- Epigenetics and DNA Methylation
Peking University
2022-2025
Capital Medical University
2024
Chengdu University of Technology
2024
Beijing Children’s Hospital
2024
Shanxi Medical University
2019-2024
Chinese Academy of Sciences
2012-2023
Peking University Stomatological Hospital
2023
Tibet University
2023
Tianjin University
2022-2023
Shanghai Institute of Organic Chemistry
2015-2023
Embryonic stem cells (ESCs) of mice and humans have distinct molecular biological characteristics, raising the question whether an earlier, "naive" state pluripotency may exist in humans. Here we took a systematic approach to identify small molecules that support self-renewal naive human ESCs based on maintenance endogenous OCT4 distal enhancer activity, signature ground pluripotency. Iterative chemical screening identified combination five kinase inhibitors induces maintains activity when...
Technologies allowing for specific regulation of endogenous genes are valuable the study gene functions and have great potential in therapeutics. We created CRISPR-on system, a two-component transcriptional activator consisting nuclease-dead Cas9 (dCas9) protein fused with activation domain single guide RNAs (sgRNAs) complementary sequence to promoters. demonstrate that can efficiently activate exogenous reporter both human mouse cells tunable manner. In addition, we show robust vivo be...
Targeted integration of transgenes can be achieved by strategies based on homologous recombination (HR), microhomology-mediated end joining (MMEJ) or non-homologous (NHEJ). The more generally used HR is inefficient for achieving gene in animal embryos and tissues, because it occurs only during cell division, although MMEJ NHEJ elevate the efficiency some systems. Here we devise a homology-mediated (HMEJ)-based strategy, using CRISPR/Cas9-mediated cleavage both transgene donor vector that...
Stimulation of TNFR1 by TNFα can promote three distinct alternative mechanisms cell death: necroptosis, RIPK1-independent and -dependent apoptosis. How cells decide which way to die is unclear. Here, we report that TNFα-induced phosphorylation RIPK1 in the intermediate domain TAK1 plays a key role regulating this critical decision. Using phospho-Ser321 as marker, show transient induced leads apoptosis when NF-κB activation inhibited cycloheximide. On other hand, blocking Ser321 promotes its...
The CRISPR/Cas9 system is an efficient gene-editing method, but the majority of gene-edited animals showed mosaicism, with editing occurring only in a portion cells. Here we show that single gene or multiple genes can be completely knocked out mouse and monkey embryos by zygotic injection Cas9 mRNA adjacent single-guide RNAs (spaced 10-200 bp apart) target key exon each gene. Phenotypic analysis F0 mice following targeted deletion eight on Y chromosome individually demonstrated robustness...
The type V-F CRISPR-Cas12f system is a strong candidate for therapeutic applications due to the compact size of Cas12f proteins. In this work, we identify six uncharacterized Cas12f1 proteins with nuclease activity in mammalian cells from assembled bacterial genomes. Among them, OsCas12f1 (433 aa) Oscillibacter sp. and RhCas12f1 (415 Ruminiclostridium herbifermentans, which respectively target 5' T-rich Protospacer Adjacent Motifs (PAMs) C-rich PAMs, show highest editing activity. Through...
The CRISPR/Cas9 system has become an efficient gene editing method for generating cells carrying precise mutations, including the rearrangement and deletion of chromosomal segments. However, whether entire chromosome could be eliminated by this technology is still unknown. Here we demonstrate use to eliminate targeted chromosomes. Using either multiple cleavages induced a single-guide RNA (sgRNA) that targets chromosome-specific sites or cocktail sgRNAs, each targeting one specific site,...
(Cell Stem Cell 15, 471–487; October 2, 2014) We have identified two errors in the data presentation version of this paper originally published online on July 24, 2014. Neither them affects results or conclusions presented any significant way. The issues are as follows: Figure 3. chemical structure shown 3C did not represent WH-4-023, but instead a closely related LCK/SRC inhibitor called WH-4-025. To correct figure we replaced with for WH-4-023 and, completeness, added citation to legend...
Regulation of ubiquitin-proteasome system (UPS), which controls the turnover short-lived proteins in eukaryotic cells, is critical maintaining cellular proteostasis. Here we show that USP14, a major deubiquitinating enzyme regulates UPS, substrate Akt, serine/threonine-specific protein kinase mediating intracellular signaling transducer for growth factors. We report Akt-mediated phosphorylation USP14 at Ser432, normally blocks its catalytic site inactive conformation, activates activity...
Current DNA base editors contain nuclease and deaminase that enables deamination of cytosine (C) or adenine (A), but no method for guanine (G) thymine (T) editing is available at present. Here we developed a deaminase-free glycosylase-based editor (gGBE) with G ability, by fusing Cas9 nickase engineered N-methylpurine glycosylase protein (MPG). By several rounds MPG mutagenesis via unbiased rational screening using an intron-split EGFP reporter, demonstrated gGBE could increase efficiency...
DNA base editors enable direct editing of adenine (A), cytosine (C), or guanine (G), but there is no editor for thymine (T) currently. Here we develop two deaminase-free glycosylase-based T (gTBE) and C (gCBE) by fusing Cas9 nickase (nCas9) with engineered human uracil glycosylase (UNG) variants. By several rounds structure-informed rational mutagenesis on UNG in cultured cells, obtain gTBE gCBE high activity T-to-S (i.e., T-to-C T-to-G) C-to-G conversions, respectively. Furthermore, conduct...
ABSTRACT Identification and the time since deposition (TsD) estimation of body fluid stains from a crime scene could provide valuable information for solving cases are always difficult forensics. Microbial characteristics were considered as promising biomarker to address issues. However, changes in microbiota may damage specific fluids. Correspondingly, incorrect identification result inaccurate TsD estimation. The mutual influence is not well understood limited codetection. In current...
<h2>Abstract</h2> Precisely targeted genome editing is highly desired for clinical applications. However, the widely used homology-directed repair (HDR)-based strategies remain inefficient certain <i>in vivo</i> We here demonstrate a microhomology-mediated end-joining (MMEJ)-based strategy precisely gene integration in transfected neurons and hepatocytes with efficiencies up to 20%, much higher (up 10 fold) than HDR-based adult mouse tissues. As proof of concept its therapeutic potential, we...
Abstract RIPK1 is a death-domain (DD) containing kinase involved in regulating apoptosis, necroptosis and inflammation. activation known to be regulated by its DD-mediated interaction ubiquitination, though underlying mechanisms remain incompletely understood. Here we show that K627 human RIPK1-DD equivalent K612 murine key ubiquitination site regulates the overall pattern of interactions with other DD-containing proteins. K627R/K612R mutation inhibits blocks both apoptosis mediated TNFR1...