CELL
·
2025
·
IF 42.5
Chao Sun,Hongchao Li,Yijing Liu,Yunjia Li,Rui Gao,Xiaoli Shi,Hongyuan Fei,Jinxing Liu,Ronghong Liang,Caixia Gao
Genome editing technologies face challenges in achieving precise, large-scale DNA manipulations in higher organisms, including inefficiency, limited editing scales and types, and the retention of undesired sequences such as recombination sites (“scars”). Here, we present programmable chromosome engineering (PCE) and RePCE, two programmable chromosome editing systems enabling scarless kilobase-to-megabase DNA manipulations in plants and human cells. Through high-throughput engineering, we obtained Lox sites with a 10-fold reduced reversibility and applied an AI-assisted recombinase engineering method (AiCE rec ) to generate Cre variants with 3.5 times the recombination efficiency of the wild type. Incorporation of a Re-pegRNA-mediated scar-free strategy further enhanced editing precision, allowing scarless insertions, deletions, replacements, inversions, and translocations at the chromosomal level. Key applications include a 315-kb inversion in rice conferring herbicide resistance, scarless chromosome fusions, and a 12-Mb inversion at human disease-related sites. These advances significantly broaden the scope of genome editing applications in molecular breeding, therapeutic development, and synthetic biology.
研究领域:
遗传学
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合成生物学
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分子生物学
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植物生物技术
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人类遗传学
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基因组编辑
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PubMed
高影响因子
TOXICOLOGY LETTERS
·
2023
·
IF 3.5
Jingjing Qu,Yinjing Zhang,Chunxia Liu,Qianqian Xie,Tong Ou,Bolin Fan,Yan Song
Anthraquinone is a recently identified contaminant present in teas globally, and its potential teratogenic and genotoxic impacts have yet to be fully comprehended. Hence, this study's objective was to determine anthraquinone's genotoxicity using various studies such as the Ames test , Mammalian erythrocyte micronucleus test, and in-vitro mammalian chromosome aberration study. Additionally, the study assessed its effects on maternal gestational toxicity and the fetus's teratogenicity through prenatal developmental toxicity research in rats. Results indicated that anthraquinone did not manifest mutagenic effects on Salmonella typhimurium histidine-deficient, did not cause chromosomal aberrations in Chinese hamster ovary cell subclone CHO-K1, and did not exhibit a genotoxic effect on mouse bone marrow erythrocytes. However, in the prenatal developmental toxicity study, administering anthraquinone orally to pregnant rats from day 5 to day 19 of gestation resulted in decreased body weight and food consumption of pregnant rats, along with a higher number of visceral malformations in the fetuses in the highest dose group (217.6 mg/kg BW). Additionally, two pregnant rats died in this group. The study has established the no observed adverse effect level (NOAEL) as 21.76 mg/kg BW, while the lowest observed adverse effect level (LOAEL) was 217.6 mg/kg BW.
研究领域:
毒理学
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遗传学
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生殖医学
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DOI
PubMed
高影响因子
JOURNAL OF APPLIED TOXICOLOGY
·
2019
·Vol. 39
·期 6
·908-918
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IF 3.065
Xiujuan Wang,Tingzhu Li,Xuerong Su,Jiangyan Li,Wenhua Li,Junying Gan,Tianshu Wu,Lu Kong,Ting Zhang,Meng Tang,Yuying Xue
With the rapid expansion of human exposure to silver nanoparticles (AgNPs), the genotoxicity screening is critical to the biosafety evaluation of nanosilver. This study assessed DNA damage and chromosomal aberration in the human hepatoma cell line (HepG2) as well as the effects on the micronucleus of bone marrow in mice induced by 20 nm polyvinylpyrrolidone-coated nanosilver (PVP-AgNPs) and 20 nm bare nanosilver (AgNPs). Our results showed that the two types of AgNPs, in doses of 20-160 μg/mL, could cause genetic toxicological changes on HepG2 cells. The DNA damage degree of HepG2 cells in 20 nm AgNPs was higher than that in 20 nm PVP-AgNPs, while the 20 nm PVP-AgNPs caused more serious chromosomal aberration than 20 nm AgNPs. Both kinds of AgNPs caused genetic toxicity in a dose-dependent manner in HepG2 cells. In the micronucleus test on mouse bone marrow cells, in doses of 10, 50 and 250 mg/kg body weight administered orally for 28 days once a day, the two kinds of AgNPs have no obvious inhibitory effect on the mouse bone marrow cells, and the effect of chromosome aberration could be documented at the high dose of 250 mg/kg. These results suggest that AgNPs have genotoxic effects in HepG2 cells and limited effects on bone marrow in mice; both in vitro and in vivo tests could be of great importance on the evaluation of genotoxicity of nanosilver. These findings can provide useful toxicological information that can help to assess genetic toxicity of nanosilver in vitro and in vivo.
研究领域:
毒理学
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遗传学
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纳米生物学
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PubMed
高影响因子
MOLECULAR BIOLOGY AND EVOLUTION
·
2022
·Vol. 39
·期 9
·
IF 8.8
Lu Meng,Li Zhi,Zhu Zi-Yu,Peng Fang,Wang Yang,Li Xi-Yin,Wang Zhong-Wei,Zhang Xiao-Juan,Zhou Li,Gui Jian-Fang
Unisexual animals are commonly found in some polyploid species complexes, and most of these species have had a long evolutionary history. However, their method for avoiding genomic decay remains unclear. The polyploid Carassius complex naturally comprises the sexual amphidiploid C. auratus (crucian carp or goldfish) (AABB) and the gynogenetic amphitriploid C. gibelio (gibel carp) (AAABBB). Recently, we developed a fertile synthetic amphitetraploid (AAAABBBB) male from C. gibelio by incorporating a C. auratus genome. In this study, we generated novel amphitriploids (AAABBB) by backcrossing the amphitetraploid male with the amphidiploid C. auratus. Whole-genome resequencing revealed the genomic changes, including recombination and independent assortment between homologs of C. gibelio and C. auratus. The fertility, sex determination system, oocyte development, and fertilization behaviors of the novel amphitriploids were investigated. Approximately 80% of the novel amphitriploid females recovered the unisexual gynogenesis ability. Intriguingly, two types of primary oocyte (with and without homolog synapsis) were discovered, and their distinct development fates were observed. Type I oocytes entered apoptosis due to improper synaptonemal complex assembly and incomplete double-strand break repair, whereas subsequent type II oocytes bypassed meiosis through an alternative ameiotic pathway to develop into mature eggs. Moreover, gynogenesis was stabilized in their offspring, and a new array of diverse gynogenetic amphitriploid clones was produced. These revealed genomic changes and detailed cytological data provide comprehensive evidence that changes in ploidy drive unisexual and sexual reproduction transition, thereby resulting in genomic diversity and allowing C. gibelio avoid genomic decay.
研究领域:
遗传学
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进化生物学
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鱼类学
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DOI
高影响因子
REGULATORY TOXICOLOGY AND PHARMACOLOGY
·
2015
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IF 2.031
Chenzhong Fei,Jie Zhang,Yang Lin,Xiaoyang Wang,Keyu Zhang,Lifang Zhang,Wenli Zheng,Mi Wang,Tao Li,Sui Xiao,Feiqun Xue,Chunmei Wang
Nitromezuril (NZL) is a novel triazine compound that exhibits remarkable anticoccidial activity. However, mutagenicity and genotoxicity of NZL have not been evaluated to date. This study evaluated the potential risks of NZL by testing for bacterial reverse mutation (Ames), mouse sperm abnormality (SA), bone marrow micronucleus (MN) and chromosomal aberration (CA). Mice were orally administered with NZL at 385, 192 and 96 mg/kg, corresponding to 0.5×, 0.25× and 0.125× the LD 50 of NZL, respectively. No significant increases in SA and CA were found in mice treated with NZL for 5 d and 3 d, respectively ( P > 0.05). NZL at 96–385 mg/kg did not have significant influence on micronucleated polychromatic erythrocyte counts ( P > 0.05). These results suggest that NZL is not genotoxic. However, Ames test results were positive both with and without the S9 system for Salmonella typhimurium TA98 and TA100, suggesting that NZL may be mutagenic. The mutagenic effects of NZL were different in in vitro and in vivo assays . Further studies should be conducted to confirm the safety of using and developing NZL as a novel anticoccidial drug.
研究领域:
毒理学
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遗传学
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PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
·
2024
·
IF 6.2
Yiren Xiong,Zuqing Hu,Di Ouyang,Meilin Tang,Jiayi He,Shanshan He,Renyi Liu,Zhenjie Gao,Ying Chen,Dalin Hu
Lead is a widespread environmental pollutant with serious adverse effects on human health, but the mechanism underlying its toxicity remains elusive. This study aimed to investigate the role of miR-584–5p / Ykt6 axis in the toxic effect of lead on HK-2 cells and the related mechanism. Our data suggested that lead exposure caused significant cytotoxicity, DNA and chromosome damage to HK-2 cells. Mechanistically, lead exposure down-regulated miR-584–5p and up-regulated Ykt6 expression, consequently, autophagosomal number and autophagic flux increased, lysosomal number and activity decreased, exosomal secretion increased. Interestingly, when miR-584–5p level was enhanced with mimic, autophagosomal number and autophagic flux decreased, lysosomal number and activity increased, ultimately, exosomal secretion was down-regulated, which resulted in significant aggravated toxic effects of lead. Further, directly blocking exosomal secretion with inhibitor GW4869 also resulted in exacerbated toxic effects of lead. Herein, we conclude that miR-584–5p / Ykt6 - mediated autophagy - lysosome - exosome pathway may be a critical route affecting the toxic effects of lead on HK-2 cells. We provide a novel insight into the mechanism underlying the toxicity of lead on human cells.
研究领域:
毒理学
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细胞生物学
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分子生物学
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DOI
PubMed
高影响因子
Antioxidants
·
2021
·Vol. 11
·期 1
·38
·
IF 6.313
Zhiwei Qiu,Jinzhao He,Guangying Shao,Jiaqi Hu,Xiaowei Li,Hong Zhou,Min Li,Baoxue Yang
Autosomal dominant polycystic kidney disease (ADPKD) is a common inherited disease characterized by progressive enlargement of fluid-filled cysts derived from renal tubular epithelial cells, which has become the fourth leading cause of end-stage renal diseases. Currently, treatment options for ADPKD remain limited. The purpose of this study was to discover an effective therapeutic drug for ADPKD. With virtual screening, Madin-Darby canine kidney (MDCK) cyst model, embryonic kidney cyst model and kidney-specificPkd1knockout mouse (PKD) model, we identified obacunone as a candidate compound for ADPKD drug discovery from a natural antioxidant compound library.In vitroexperiments showed that obacunone significantly inhibited cyst formation and expansion of MDCK cysts and embryonic kidney cysts in a dose-dependent manner.In vivo, obacunone treatment significantly reduced the renal cyst development in PKD mice. Western blot and morphological analysis revealed that obacunone served as a NRF2 activator in ADPKD, which suppressed lipid peroxidation by up-regulating GPX4 and finally restrained excessive cell proliferation by down-regulating mTOR and MAPK signaling pathways. Experimental data demonstrated obacunone as an effective renal cyst inhibitor for ADPKD, indicating that obacunone might be developed into a therapeutic drug for ADPKD treatment.Keywords:obacunone;autosomal dominant polycystic kidney disease;NRF2;glutathione peroxidase;lipid peroxidation;MAPK;mTOR
研究领域:
药理学
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肾病学
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分子生物学
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PubMed
高影响因子
Nature Communications
·
2025
·Vol. 16
·期 1
·1-20
·
IF 15.7
Phillips Aaron F.,Zhang Rumin,Jaffe Mia,Schulz Ryan,Carty Marysol Chu,Verma Akanksha,Feinberg Tamar Y.,Arensman Michael D.,Chiu Alan,Letso Reka,Bosco Nazario,Queen Katelyn A.,Racela Allison R.,Stumpff Jason,Andreu-Agullo Celia,Bettigole Sarah E.,Depetris Rafael S.,Drutman Scott,Su Shinsan M.,Cogan Derek A.,Eng Christina H.
Chromosome instability is a prevalent vulnerability of cancer cells that has yet to be fully exploited therapeutically. To identify genes uniquely essential to chromosomally unstable cells, we mined the Cancer Dependency Map for genes essential in tumor cells with high levels of copy number aberrations. We identify and validate KIF18A, a mitotic kinesin, as a vulnerability of chromosomally unstable cancer cells. Knockdown of KIF18A leads to mitotic defects and reduction of tumor growth. Screening of a chemical library for inhibitors of KIF18A enzymatic activity identified a hit that was optimized to yield VLS-1272, which is orally bioavailable, potent, ATP non-competitive, microtubule-dependent, and highly selective for KIF18A versus other kinesins. Inhibition of KIF18A’s ATPase activity prevents KIF18A translocation across the mitotic spindle, resulting in chromosome congression defects, mitotic cell accumulation, and cell death. Profiling VLS-1272 across >100 cancer cell lines demonstrates that the specificity towards cancer cells with chromosome instability differentiates KIF18A inhibition from other clinically tested anti-mitotic drugs. Treatment of tumor xenografts with VLS-1272 results in mitotic defects leading to substantial, dose-dependent inhibition of tumor growth. The strong biological rationale, robust preclinical data, and optimized compound properties enable the clinical development of a KIF18A inhibitor in cancers with high chromosomal instability.
研究领域:
肿瘤学
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分子生物学
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遗传学
·
药物研发
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PubMed
高影响因子
TOXICOLOGY IN VITRO
·
2017
·
IF 2.866
Y.F. Zhang,S.Y. Chen,M.J. Qu,A.O. Adeleye,Y.N. Di
Freshly isolated cells are used as an ideal experimental model in in vitro toxicology analysis, especially the detection of diverse xenobiotics induced genotoxic effects. In present study, heavy metals (Zn, Cu, Cd, Pb) and PCBs were selected as representative xenobiotics to verify the ability of in vitro model in assessing genotoxic effects in cells of marine mussels ( Mytilus galloprovincialis ). DNA damage and chromosome aberration were assessed in freshly isolated cells from haemolymph, gill and digestive gland by single cell gel electrophoresis and micronucleus assay respectively. Gill cells showed more sensitive to Zn exposure among three types of cells, indicating tissue-specific genotoxicity. Significantly higher DNA aberrations were induced by Cu in haemocytes compared to Cd and Pb, indicating chemical-specific genotoxicity. An additive effect was detected after combined heavy metals and PCBs exposure, suggesting the interaction of selected xenobiotics. To our knowledge, this is the first attempt to study the complex effects of organic and/or inorganic contaminants using freshly isolated cells from marine mussels. Genetic responses are proved to occur and maintained in vitro in relation to short-term xenobiotics induced stresses. The utilization of the in vitro model could provide a rapid tool to investigate the comprehensive toxic effects in marine invertebrates and monitor environmental health.
研究领域:
海洋毒理学
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环境科学
·
分子生物学
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DOI
PubMed
高影响因子
TOXICOLOGY
·
2024
·
IF 4.8
Yiren Xiong,Guoqiang Guo,Hongyi Xian,Zuqing Hu,Di Ouyang,Jiayi He,Shanshan He,Renyi Liu,Zhenjie Gao,Meilin Tang,Ying Chen,Suqin Tan,Xiaoqi Zhu,Abudumijiti Abulimiti,Sujin Zheng,Hehai Huang,Dalin Hu
Tributyltin chloride (TBTC) is a ubiquitous environmental pollutant with various adverse effects on human health. Exosomes are cell - derived signaling and substance transport vesicles. This investigation aimed to explore whether exosomes could impact the toxic effects caused by TBTC via their transport function. Cytotoxicity, DNA and chromosome damage caused by TBTC on MCF-7 cells were analyzed with CCK-8, flow cytometry, comet assay and micronucleus tests, respectively. Exosomal characterization and quantitative analysis were performed with ultracentrifugation, transmission electron microscope (TEM) and bicinchoninic acid (BCA) methods. TBTC content in exosomes was detected with Liquid Chromatography-Mass Spectrometry (LC-MS). The impacts of exosomal secretion on the toxic effects of TBTC were analyzed. Our data indicated that TBTC caused significant cytotoxicity, DNA and chromosome damage effects on MCF-7 cells, and a significantly increased exosomal secretion. Importantly, TBTC could be transported out of MCF-7 cells by exosomes. Further, when exosomal secretion was blocked with GW4869, the toxic effects of TBTC were significantly exacerbated. We concluded that TBTC promoted exosomal secretion, which in turn transported TBTC out of the source cells to alleviate its toxic effects. This investigation provided a novel insight into the role and mechanism of exosomal release under TBTC stress.
研究领域:
毒理学
·
细胞生物学
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环境科学
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药理学
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DOI
PubMed
高影响因子
EUPHYTICA
·
2024
·Vol. 220
·期 4
·1-13
·
IF 1.6
Zhong Yuan,Du Ming-jie,Ji Run-ze,Rong Qi,Guan Zi-heng,Cheng Fang-yun
Cross-incompatibility is the major challenge in inter-sectional distant hybridization between tree peonies ( Paeonia sect. Moutan ) and herbaceous peonies ( Paeonia sect. Paeonia ). Most of the intersectional cross combinations rarely produce hybrid progenies or even seeds. However, a combination with Paeonia × lemoinei Rehd. ( P. delavayi × P. suffruticosa , tree type) as the pollen parent and P. lactiflora (herbaceous) as the seed parent resulted in many progenies. This research focuses on cytogenetics of two highly compatible intersectional combinations, P. lactiflora ‘Martha W.’ × P. × lemoinei ‘Golden Era’ (‘MW’ × ‘GE’) and P. lactiflora ‘Fen Yun Fei He’ × P. × lemoinei ‘L’Esperance’ (‘FYFH’ × ‘LE’). Karyotype analysis combined with genomic in situ hybridization (GISH) showed that the seed parents, ‘MW’ and ‘FYFH’, are both diploids (2 n = 2 x = 10, CC), while the pollen parents, ‘GE’ and ‘LE’, are tetraploid (2 n = 4 x = 20, AABB) and diploid (2 n = 2 x = 10, AB) respectively. Four progenies from ‘MW’ × ‘GE’, and another four progenies from ‘FYFH’ × ‘LE’, are all triploids (2 n = 3 x = 15, ABC). P. × lemoinei contributes two sets of chromosomes (AB) showing distinct signals in all tested triploids. The essential of a successful intersectional hybridization is to obtain 2 x gametes from tree peony. P. × lemoinei has better compatibility in intersectional hybridization than other tree peony species, probably because of its higher frequency of 2 x gamete (unreduced in diploid, or reduced in tetraploid) generation. Tetraploid tree peonies, such as ‘GE’, might produce abundant 2 x gametes through normal meiosis, which is the possible reason for their better fertility and superior intersectional compatibility than diploid tree peonies. The mechanism of compatibility in intersectional hybridization is supposed to be a rebalance between the distant divergent parents, tree peonies and herbaceous peonies, triggered by ploidy increase of gametes from tree peonies. The discovery of tetraploid tree peony also provides valuable materials to study the mechanism of polyploidization in sect. Moutan , and to promote the distant hybridization breeding in Paeonia.
研究领域:
植物学
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遗传学
·
植物育种
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DOI
高影响因子
Stem Cell Research & Therapy
·
2017
·Vol. 8
·期 1
·1-13
·
IF 4.211
Wang Linli,Chen Yuehua,Guan Chunyan,Zhao Zhiju,Li Qiang,Yang Jianguo,Mo Jian,Wang Bin,Wu Wei,Yang Xiaohui,Song Libing,Li Jun
Background Because the lack of an induced pluripotent stem cell (iPSC) induction system with optimal safety and efficiency limits the application of these cells, development of such a system is important.Methods To create such an induction system, we screened a variety of reprogrammed plasmid combinations and multiple compounds and then verified the system’s feasibility using urine cells from different individuals. We also compared large-scale iPSC chromosomal variations and expression of genes associated with genomic stability between this system and the traditional episomal system using karyotype and quantitative reverse transcription polymerase chain reaction analyses.Results We developed a high-efficiency episomal system, the 6F/BM1-4C system, lacking tumorigenic factors for human urine-derived cell (hUC) reprogramming. This system includes six low-risk factors (6F), Oct4, Glis1, Klf4, Sox2, L-Myc, and the miR-302 cluster. Transfected hUCs were treated with four compounds (4C), inhibitor of lysine-demethylase1, methyl ethyl ketone, glycogen synthase kinase 3 beta, and histone deacetylase, within a short time period. Comparative analysis revealed significantly decreased chromosomal variation in iPSCs and significantly increased Sirt1 expression compared with iPSCs induced using the traditional episomal system.Conclusion The 6F/BM1-4C system effectively induces reprogramming of urine cells in samples obtained from different individuals. iPSCs induced using the 6F/BM1-4C system are more stable at the cytogenetic level and have potential value for clinical application.
研究领域:
干细胞生物学
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基因组学
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医学研究
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PubMed
高影响因子
危险品化学品经营许可证（带存储）