Tumor necrosis factor-stimulated gene-6 ameliorates early brain injury after subarachnoid hemorrhage by suppressing NLRC4 inflammasome-mediated astrocyte pyroptosis

Subarachnoid hemorrhage is associated with high morbidity and mortality and lacks effective treatment.Pyroptosis is a crucial mechanism underlying early brain injury after subarachnoid hemorrhage.Previous studies have confirmed that tumor necrosis factor-stimulated gene-6(TSG-6)can exert a neuroprotective effect by suppressing oxidative stress and apoptosis.However,no study to date has explored whether TSG-6 can alleviate pyroptosis in early brain injury after subarachnoid hemorrhage.In this study,a C57BL/6J mouse model of subarachnoid hemorrhage was established using the endovascular perforation method.Our results indicated that TSG-6 expression was predominantly detected in astrocytes,along with NLRC4 and gasdermin-D(GSDMD).The expression of NLRC4,GSDMD and its N-terminal domain(GSDMD-N),and cleaved caspase-1 was significantly enhanced after subarachnoid hemorrhage and accompanied by brain edema and neurological impairment.To explore how TSG-6 affects pyroptosis during early brain injury after subarachnoid hemorrhage,recombinant human TSG-6 or a siRNA targeting TSG-6 was injected into the cerebral ventricles.Exogenous TSG-6 administration downregulated the expression of NLRC4 and pyroptosis-associated proteins and alleviated brain edema and neurological deficits.Moreover,TSG-6 knockdown further increased the expression of NLRC4,which was accompanied by more severe astrocyte pyroptosis.In summary,our study revealed that TSG-6 provides neuroprotection against early brain injury after subarachnoid hemorrhage by suppressing NLRC4 inflammasome activation-induced astrocyte pyroptosis.

16S rRNA Gene-Based Metagenomic Analysis of Soil Bacterial Diversity in Brazzaville, Republic of the Congo

Soil contains a great diversity of microorganisms, among which are bacteria. This study aimed to explore bacterial diversity in soil samples in Brazzaville in the Republic of the Congo. Environmental DNA was extracted. The illumina MiSeq sequencing was held and the diversity indices have been computed. Illumina MiSeq sequencing revealed 21 Phyla, four of which were abundant: Proteobacteria, Acidobacteria, Actinobacteria and Bacteroidetes. Soil microbial communities in the studied samples were phylogenetically diverse but with a stable community structure. 17 classes are represented with relative abundances of Rihzobiales, Bacillales, Actinomycetales and Acidobacteriales. 40 families, the Alphaproteobacteria, the Bacilli and the 12 Actinobacteria. 83 orders among which the Rhizobiales are the most abundant followed by Bacillales and the least abundant followed by the Flavobacteriaceae. Of the 28 genera listed, the Bradyrhizobium is the most dominant in Mw3 and Mw4. 25 listed species, Bradyrhizobium, Bacillus, Actinoplanes, and Candidatu coribacter Acidobacterium are the most abundant species. The Shannon indices of Mw3 and Mw4 are equal, the H’max of Mw4 is greater than the H’max of Mw3. The Simpson index of Mw4 is equal to the Simpson index of Mw3, and the Pielou index (J) of Mw4 is less than the R of Mw3, but very close. This study opens interesting perspectives on the knowledge and exploitation of telluric bacteria in several areas of life.

Gene-Xpert MTB/RIF在西藏地区结核病诊断中的应用研究

目的探讨Gene-Xpert MTB/RIF在西藏地区结核病诊断中的应用价值。方法对974例可疑结核病患者的标本同时进行结核涂片抗酸染色镜检和Gene-Xpert MTB/RIF检测。对比两种检测结果。结果 974例检测标本中,Gene-Xpert MTB/RIF检测阳性率为12%(118/974),其中利福平敏感的标本为93例,利福平耐药的标本为25例(耐药率为21%)。抗酸染色阳性标本为15例,检测阳性率为2%。Gene-Xpert MTB/RIF检测阳性率高于抗酸染色,差异具有统计学意义(P<0.05)。结论 Gene-Xpert MTB/RIF在西藏地区结核病诊断和治疗中具有较好的临床应用价值,且方法简便、快速。

Gene-gene,gene-environment,gene-nutrient interactions and single nucleotide polymorphisms of inflammatory cytokines

Inflammation plays a significant role in the etiology of type 2 diabetes mellitus(T2DM).The rise in the pro-inflammatory cytokines is the essential step in glucotoxicity and lipotoxicity induced mitochondrial injury,oxidative stress and beta cell apoptosis in T2 DM.Among the recognized markers are interleukin(IL)-6,IL-1,IL-10,IL-18,tissue necrosis factor-alpha(TNF-α),C-reactive protein,resistin,adiponectin,tissue plasminogen activator,fibrinogen and heptoglobins.Diabetes mellitus has firm genetic and very strong environmental influence; exhibiting a polygenic mode of inheritance.Many single nucleotide polymorphisms(SNPs) in various genes including those of pro and antiinflammatory cytokines have been reported as a risk for T2 DM.Not all the SNPs have been confirmed by unifying results in different studies and wide variations have been reported in various ethnic groups.The inter-ethnic variations can be explained by the fact that gene expression may be regulated by gene-gene,gene-environment and gene-nutrient interactions.This review highlights the impact of these interactions on determining the role of single nucleotide polymorphism of IL-6,TNF-α,resistin and adiponectin in pathogenesis of T2 DM.

＂外源基因清除＂技术(Gene-Deletor)原理、特点及其潜在应用前景

"外源基因清除"技术(Gene-Deletor)是美国康涅狄格大学华裔教授李义领导的研究小组发展起来的,有望成为解决转基因植物环境安全性和食品安全性问题的有力工具。象计算机"卸载"应用软件一样,"外源基因清除"技术利用器官特异或诱导型启动子、重组酶及融合识别位点构建基因表达元件,能将转基因植物中的全部外源基因在完成其功能作用后自动地从花粉、种子和果实中彻底清除。最近,美国著名"社会技术"公司公布了未来20年对社会最具影响力、最有前景的技术创新领域一系列12个简报,其中第10个专题推介了"外源基因清除"技术。本文综述了"外源基因清除"技术的原理和技术优点,介绍并讨论了李义对该技术在作物有性繁殖中的应用以及在解决食品安全性问题上的应用方略。

基于GENE-8310的嵌入式TinyOs系统设计

无线传感器网络是当前国际上备受关注的、多学科高度交叉、知识高度集成的前沿热点研究技术,其核心技术Tinyos被誉为是"无线嵌入式系统"。在嵌入式开发板GENE-8310上移植Tinyos应用操作系统是一次技术上的新尝试,将GENE-8310作为无线传感器网络中的远程服务器,实现无线网络与有线网络的跨网段传输和远程网络监控将进一步推动无线传感器网络的技术的发展。

siRNA下调astrocyte elevated gene-1对神经母细胞瘤细胞增殖和凋亡影响的体外研究

目的:探讨siRNA下调astrocyte elevated gene-1(AEG-1)表达对神经母细胞瘤细胞增殖、凋亡和细胞周期的影响。方法:用设计合成的AEG-1 siRNA转染神经母细胞瘤细胞株M17和SK-N-SH,采用荧光定量RT-PCR技术观察AEG-1 siRNA对AEG-1基因表达的影响;用MTT法和克隆形成实验观察AEG-1 siRNA对神经母细胞瘤细胞的增殖抑制作用;用流式细胞术检测下调AEG-1表达对神经母细胞瘤细胞凋亡及细胞周期的影响。结果:用AEG-1 siRNA转染人神经母细胞瘤细胞,RT-PCR结果证实AEG-1 siRNA能显著基因下调AEG-1表达,与对照组相比有显著差异(P<0.01);MTT法和克隆形成实验结果证明下调AEG-1表达可显著抑制神经母细胞瘤细胞的增殖;流式细胞术检测结果表明下调AEG-1表达可促进神经母细胞瘤细胞凋亡,并将细胞周期阻滞在G_0/G_1期。结论:AEG-1 siRNA可下调基因在神经母细胞瘤细胞中表达,并抑制神经母细胞瘤细胞增殖,促进细胞凋亡。

Astrocyte elevated gene-1的研究进展

Astrocyte elevated gene-1 (AEG-1) was cloned as an human immunodeficiency virus -1-inducible and tumor necrosis factor-α-inducible transcript in primary human fetal astrocytes by a rapid subtraction hybridization approach. AEG-1 down-regulates the expression of the glutamate transporter EAAT2,thus,it is implicated in glutamate-induced excitotoxic damage to neurons as evident in HIV-associated neurodegeneration. Meanwhile,AEG-1 expression is elevated in subsets of breast cancer,prostatic cancer,glioblastoma multiforme and melanoma cells,having a dual specificity phosphatase activity. Overexpression of AEG-1 increases and siRNA inhibition of AEG-1 decreases migration and invasion of human glioma cells,respectively. Recent observations indicate that AEG-1 exerts its effects by activating the nuclear factor kappa B (NF-κB) pathway and AEG-1 is a downstream target of Ha-ras and plays an important role in Ha-ras-mediated tumorigenesis. These findings are intensifying interest in AEG-1 as a crucial regulator of tumor progression and metastasis and as a potential mediator of neurodegeneration.

Lipofectamine^TM和Fugene-6介导GFP基因转染绵羊胎儿成纤维细胞的比较研究

分别用LipofectamineTM和Fegene-6介导质粒pEGFP-C1转染体外培养的绵羊胎儿成纤维细胞(sheepfetal fibroblast cells,sFFCs),比较了DNA浓度、转染试剂用量以及细胞暴露于DNA、转染试剂中作用时间对转染效率的影响,通过含G418的DMEM/F12培养液筛选得到转基因单克隆细胞。结果表明脂质体转染试剂LipofectamineTM转染效率优于Fegene-6。另外,对转基因细胞进行了染色体核型分析。结果表明,转基因细胞中二倍体核型占74.5%,与对照组比较没有显著性差异。以上研究为其他基因转染sFFCs以及利用体细胞克隆法生产转基因绵羊提供了参考依据。

DNA-微阵列芯片检测技术与Gene-Xpert MTB/RIF技术在新疆维吾尔自治区结核分枝杆菌耐药性诊断中的应用

目的探索DNA-微阵列芯片检测技术(以下简称基因芯片技术)和利福平耐药实时荧光定量核酸扩增检测技术(Gene-Xpert MTB/RIF技术)检测新疆维吾尔自治区结核分枝杆菌中利福平和异烟肼耐药性的应用价值。方法收集2015年新疆维吾尔自治区结核病耐药监测菌株115株,对所有的菌株进行比例法药敏试验、基因芯片技术、Gene-Xpert MTB/RIF技术检测。以比例法药敏试验为“金标准”,分别计算基因芯片技术与Gene-Xpert MTB/RIF技术对结核分枝杆菌耐药性诊断的效能并进行评价。结果比例法药敏试验结果显示,利福平耐药患者40例(34.78%),异烟肼耐药患者48例(41.74%)。以比例法药敏试验结果为“金标准”,基因芯片技术检测结核分枝杆菌对利福平和异烟肼耐药耐药性诊断的灵敏度、特异度分别为80.00%、89.33%和77.08%、97.01%,Gene-Xpert MTB/RIF技术检测结核分枝杆菌对利福平耐药性诊断的灵敏度、特异度分别为92.50%、94.67%。基因芯片技术和Gene-Xpert MTB/RIF技术对利福平耐药患者的诊断效能差异无统计学意义(P>0.05)。结论对于新疆维吾尔自治区结核病耐药监测菌株,基因芯片技术和Gene-Xpert MTB/RIF技术对耐药结核病的诊断均具有较高的灵敏度和特异度。Gene-Xpert MTB/RIF技术操作方法简单,检测时间短;基因芯片技术耐药检测范围广,耗时少,为新疆维吾尔自治区广泛开展耐药结核病快速检测提供了有力的技术支持。

Expression of hypoxia-inducible factor 1 alpha and oligodendrocyte lineage gene-1 in cultured brain slices after oxygen-glucose deprivation

Oligodendrocyte lineage gene-1 expressed in oligodendrocytes may trigger the repair of neuronal myelin impairment, and play a crucial role in myelin repair. Hypoxia-inducible factor la, a transcription factor, is of great significance in premature infants with hypoxic-ischemic brain damage There is little evidence of direct regulatory effects of hypoxia-inducible factor le on oligodendrocyte lineage gene-l. In this study, brain slices of Sprague-Dawley rats were cultured and subjected to oxygen-glucose deprivation. Then, slices were transfected with hypoxia-inducible factor la or oligodendrocyte lineage gene-1. The expression levels of hypoxia-inducible factor la and oligodendrocyte lineage gene-1 were significantly up-regulated in rat brains prior to transfection, as detected by immunohistochemical staining. Eight hours after transfection of slices with hypoxia-inducible factor la, oligodendrocyte lineage gene-1 expression was upregulated, and reached a peak 24 hours after transfection. Oligodendrocyte lineage gene-1 transfection induced no significant differences in hypoxia-inducible factor la levels in rat brain tissues with oxygen-glucose deprivation. These experimental findings indicate that hypoxia-inducible factor la can regulate oligodendrocyte lineage gene-1 expression in hypoxic brain tissue, thus repairing the neural impairment.

16S rRNA Gene-Based Analysis of Ileal Bacterial Community and Phylogeny in Nursing and Weaned Piglets

Weaning of piglets is generally considered as a stressor which changes intestinal ecosystem and leads to clinical implications. Microbiota inhabiting in small intestine （especially ileum） are assumed to promote health, but their functional properties are yet poody dascdbed. As indicated by the 16S rRNA gene sequences of ileal micrebiota in nursing piglets （at the age of 21 and 28 d） and 28-day-old weaned piglets （weaned at 21 d of age）, the microbiota were mainly comprised of gram-positive bacteria. There were 40 operational taxonomic units （OTUs） （from 171 clones） in the ileum of nursing piglets aged 21 d, 61 OTUs （from 194 clones） in the ileum of nursing piglets aged 28 d, and 56 OTUs （from 171 clones） in the ileum of weaned piglets aged 28 d. The flea of nursing piglets aged 21 d were dominantly occupied by Lactobacilli （87.7%） as well as Streptococ cus （ 3.5 % ）. Lactobacillus amy/ovorus （41.5 % ）, Lactobaci/lus sp. （ 19.3 % ）, Lactobaci/lus reuteri （ 12.3 % ）, Lactobacillus salivarius （ 9.4 % ） and L. mucosae （4.7%） were the predominant species among Lactobacil/L Similar results were obtained in the nursing piglets at 28 d of age ex- cept that Lactobaci/li decreased to 71.1% and Streptococcus increased to 21.1% significantly. Lactobacillus （52.0%） and Streptococcus （26.3%） were the two major groups in the ileum of weaned piglets aged 28 d. Lactobacillus amylovorus （31.6%） and Lactobaci/lus reuteri （ 16.4% ） was the two most important species in Lactobacillus. Therefore, Lactobacilli were predominant in the ileum of nursing and weaned piglets, and they had the highest diversity, followed by Streptococcus. The diversity of ileal microbiota was not different remarkably between the nursing piglets and the weaned piglets, but the composition changed significantly. These findings are helpful to understand ileal bacterial ecophysiology and further develop nutritional regimes to prevent or counteract complications during the weaning transition.

Downregulation of Astrocyte Elevated Gene-1 Expression Combined with All-Trans Retinoic Acid Inhibits Development of Vasculogenic Mimicry and Angiogenesis in Glioma

Objective This study aimed to investigate the effects of downregulating astrocyte elevated gene-1(AEG-1)expression combined with all-trans retinoic acid(ATRA)on vasculogenic mimicry(VM)formation and angiogenesis in glioma.Methods U87 glioma cells were transfected with AEG-1 shRNA lentiviral vectors(U87-siAEG-1)and incubated in a medium containing 20µmol/L ATRA.Matrigel-based tube formation assay was performed to evaluate VM formation,and the cell counting kit-8(CCK-8)assay was used to analyze the proliferation of glioma cells in vitro.Reverse transcription-quantitative polymerase chain reaction and Western blot analysis were used to investigate the mRNA and protein expression of related genes,respectively.Glioma xenograft models were generated via subcutaneous implantation of glioma cells in nude mice.Tumor-bearing mice received an intraperitoneal injection of ATRA(10 mg/kg per day).Immunohistochemistry was used to evaluate the expression of related genes and the microvessel density(MVD)in glioma xenograft models.CD34/periodic acid-Schiff double staining was performed to detect VM channels in vivo.The volume and weight of tumors were measured,and a tumor growth curve was drawn to evaluate tumor growth.Results A combination of ATRA intervention and downregulation of AEG-1 expression significantly inhibited the proliferation of glioma cells in vitro and glioma VM formation in vitro and in vivo.It also significantly decreased MVD and inhibited tumor growth.Further,the expression levels of matrix metalloproteinase(MMP)-2,MMP-9,vascular endothelial-cadherin(VE-cadherin),and vascular endothelial growth factor(VEGF)in glioma significantly decreased in vivo and in vivo.Conclusion Hence,a combinatorial approach might be effective in treating glioma through regulating MMP-2,MMP-9,VEGF,and VE-cadherin expression.

Enhanced Effects of TRAIL-endostatin-based Double-gene-radiotherapy on Suppressing Growth, Promoting Apoptosis and Inducing Cell Cycle Arrest in Vascular Endothelial Cells

This study examined the effects of TRAIL-endostatin-based gene-radiotherapy on cellu-lar growth, apoptosis and cell cycle progression in human vascular endothelial cells ECV304 in vitro. The expression of TRAIL and endostatin protein in ECV304 cells was detected by ELISA after the transfection of recombinant plasmid pshuttle-Egr1-shTRAIL-shES and X-ray irradiation. Then MTT assay was used for determining the cellular proliferation, and flow cytometry (FCM) plus Annexin V and propidium iodide (PI) double-staining or PI single-staining were employed for the detection of apoptosis and cell cycle progression. The results showed that expression of TRAIL and endostatin protein exhibited a time- and dose-dependent change in ECV304 cells after pshut-tle-Egr1-shTRAIL-shES transfection in conjunction with irradiation. In the TRAIL-endostatin-based single- or double-gene-radiotherapy, the cell viability declined in a time- and dose-dependent manner, the percentage of cells at G2/M phase and apoptotic rate was increased, and the percentage of cells at G0/G1 phase was lowered as compared with those receiving radiotherapy alone. Moreover, TRAIL-endostatin-based double-gene-radiotherapy demonstrated better effects on growth inhibition, promotion of apoptosis and induction of cell cycle arrest in ECV304 cells than single-gene-radiotherapy.

胸腔镜联合胸水gene-Xpert检查在结核性胸膜炎诊断中的价值

目的观察在结核性胸膜炎诊断中应用胸腔镜联合胸水gene-Xpert检查的临床价值。方法采用回顾性研究方法,选取2019年6月至2020年6月沈阳市第十人民医院呼吸科收治的结核性胸膜炎患者200例纳入本次研究,所有患者均给予常规抗结核治疗。以胸液抗酸杆菌培养为诊断金标准,分别评价胸腔镜检查、胸水gene-Xpert检查、联合检查诊断结核性胸膜炎的准确性、特异度和敏感度。结果胸腔镜下活检诊断为结核性胸膜炎有144例,占72.0%(127/200);胸水gene-Xpert检查诊断为结核性胸膜炎有143例,占71.5%(143/200);联合检查诊断为结核性胸膜炎有154例,占77.0%(127/200)。胸腔镜下活检的诊断结核性胸膜炎的准确性为72.0%,特异度为30.9%,敏感度为87.6%;胸水gene-Xpert检查的诊断结核性胸膜炎的准确性为71.5%,特异度为31.0%,敏感度为88.0%;联合检查的诊断结核性胸膜炎的准确性为77.0%,特异度为67.5%,敏感度为94.1%。联合检查的诊断结核性胸膜炎的特异度、敏感度均明显高于单独的胸腔镜下活检和胸水gene-Xpert检查,差异有统计学意义(P<0.05)。结论在结核性胸膜炎的临床诊断中,胸腔镜联合胸水gene-Xpert检查具有较高的诊断准确性和特异度,利于病情的快速、准确诊断,为患者的早期确诊、早期治疗提供保障,体现出很高的临床应用价值。

Regulative Effect of Traditional Chinese Medicine on Gene-expression Related to Precancerous Lesion of Gastric Cancer

The gene-expression changes related with precancerous lesion of gastric cancer (PLGC) are surveyed. Not only the regulative effect of traditional Chinese medicine (TCM) on oncogene, antioncogene and anti-apoptosis gene that are related with PLGC is analyzed, but also current research state is presented. It's showed that TCM has effects of therapy and inversion on PLGC. These effects are related with the inhibition to related oncogene expression, the regulation and activation to the deletion of antioncogene, the inhibition to the high-expression of mutant gene-protein about antioncogene, and the regulative function to anti-apoptosis gene.

Use of gene-editing technology to introduce targeted modifications in pigs

Pigs are an important resource in agriculture and serve as a model for human diseases. Due to their physiological and anatomical similarities with humans, pigs can recapitulate symptoms of human diseases, making them a useful model in biomedicine. However, in the past pig models have not been widely used partially because of the difficulty in genetic modification. The lack of true embryonic stem cells in pigs forced researchers to utilize genetic modification in somatic cells and somatic cell nuclear transfer(SCNT) to generate genetically engineered(GE) pigs carrying site-specific modifications. Although possible, this approach is extremely inefficient and GE pigs born through this method often presented developmental defects associated with the cloning process. Advancement in the gene-editing systems such as Zinc-Finger Nucleases(ZFNs), Transcription activator-like effector nucleases(TALENs), and the Clustered regularly interspaced short palindromic repeat(CRISPR)/CRISPR-associated 9(Cas9) system have dramatically increased the efficiency of producing GE pigs. These gene-editing systems, specifically engineered endonucleases, are based on inducing double-stranded breaks(DSBs) at a specific location, and then site-specific modifications can be introduced through one of the two DNA repair pathways: non-homologous end joining(NHEJ) or homology direct repair(HDR).Random insertions or deletions(indels) can be introduced through NHEJ and specific nucleotide sequences can be introduced through HDR, if donor DNA is provided. Use of these engineered endonucleases provides a higher success in genetic modifications, multiallelic modification of the genome, and an opportunity to introduce site-specific modifications during embryogenesis, thus bypassing the need of SCNT in GE pig production. This review will provide a historical prospective of GE pig production and examples of how the gene-editing system, led by engineered endonucleases, have improved GE pig production. We wil also present some of our current progress related to the optimal use of CRISPR/Cas9 system during embryogenesis.

Susceptibility to ulcerative colitis in Hungarian patients determined by gene-gene interactions

AIM: To study the inflammatory bowel disease-5 locus (IBD5) and interleukin-23 receptor (IL23R) gene variants in UC patients and test for gene-gene interaction.

Preliminary study on androgen dependence of calcitonin gene-related peptide in rat penis

Aim: To study the androgen dependence of the neurotransmitter, calcitonin gene-related peptide (CGRP) in rat penis. Methods: Forty-four Sprague-Dawley rats were randomly divided into Group A (intact controls), Group B (castrated) and Group C (gavaged with finasteride 4.5 mg·kg^(-1).day^(-1)). Four and ten weeks later respectively, half of rats in each group were anaesthetized. Blood samples were taken for the measurement of serum testosterone and dihydrotestosterone (DHT) by means of radioimmunoassay. Penile samples were harvested for the investigation of calcitonin gene related peptide (CGRP)-immunoreactive nerve fibers with immunohistochemistry. The computer-assisted imaging analysis system was applied to calculate the area proportion of the CGRP-positive nerve fibers (CGRP-PNF) in each group. Results: 1) Both 4 and 10 weeks later, testosterone and DHT levels in Group B decreased significantly compared with those in Group A, (P < 0.05, P < 0.01, respectively); DHT level in Group C was also significantly decreased in comparison with that in Group A for both 4- and 10- week animals (P < 0.05); 2) There was no significant differences in area proportion of CGRP-PNF among Groups A, B and C 4 weeks after treatments (P > 0.05); However, 10 weeks later, the proportion of CGRP-PNF in Groups B and C was significantly less than that in Group A (P < 0.01);3) The proportion of CGRP-PNF of 4-week animals in Groups B and C was significantly higher than that of 10-week animals (P < 0.05). Conclusion: The expression of neurotransmitter, CGRP may depend on androgens, including testosterone and DHT in rat penis.

Improvements of TKC Technology Accelerate Isolation of Transgene-Free CRISPR/Cas9-Edited Rice Plants

Elimination of the CRISPR/Cas9 constructs in edited plants is a prerequisite for assessing genetic stability, conducting phenotypic characterization, and applying for commercialization of the plants. However, removal of the CRISPR/Cas9 transgenes by genetic segregation and by backcross is laborious and time consuming. We previously reported the development of the transgene killer CRISPR(TKC) technology that uses a pair of suicide genes to trigger self-elimination of the transgenes without compromising gene editing efficiency. The TKC technology enables isolation of transgene-free CRISPR-edited plants within a single generation, greatly accelerating crop improvements. Here, we presented two new TKC vectors that show great efficiency in both editing the target gene and in undergoing self-elimination of the transgenes. The new vectors replaced the CaMV35 S promoter used in our previous TKC vector with two rice promoters to drive one of the suicide genes, providing advantages over our previous TKC vector under certain conditions. The vectors reported here offered more options and flexibility to conduct gene editing experiments in rice.