检索结果-南通市图书馆

Targeted mutagenesis in rice using CRISPR-Cpf1 system

在线全文

学校读者我要写书评

暂无评论

Journal of Genetics and Genomics 2017年第1期44卷 71-73页

作者： xixun hu Chun Wang Qing Liu Yaping Fu Kejian Wang State Key Laboratory of Rice Biology China National Rice Research InstituteChinese Academy of Agricultural Sciences Hangzhou 310006 China

Cpf1 is a class 2/type V CRISPR effector that has been recently harnessed for genome editing （Zetsche et al., 2015; hut et al., 2016; Kim et al., 2016）. Cpff recognizes thymidine-rich sequence as the protospacer-adjacent motif （PAM） at the 5＇ end of target sequences. In addition, Cpfl requires only a single shorter crRNA and cleaves DNA in a staggered fashion with 5＇ overhangs （Zetsche et al., 2015）.

关键词： Targeted mutagenesis in rice using CRISPR-Cpf1 system tRNA

Improving the efficiency of the CRISPR-Cas12a system with tRNA-crRNA arrays

在线全文

学校读者我要写书评

暂无评论

The Crop Journal 2020年第3期8卷 403-407页

作者： xixun hu Xiangbing Meng Jiayang Li Kejian Wang Hong Yu State Key Laboratory of Rice Biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 310006ZhejiangChina State Key Laboratory of Plant Genomics and National Center for Plant Gene Research(Beijing) Institute of Genetics and Developmental BiologyThe Innovative Academy of Seed DesignChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China

CRISPR-Cas12a offers a convenient tool for multiplex genome editing in rice. However, the CRISPR-Cas12a system displays variable editing efficiency across genomic loci, with marked influence by CRISPR RNAs(crRNAs). To improve the efficiency of the CRISPR-Cas12a system for multiplex genome editing, we identified various architectures and expression strategies for crRNAs. Transformation of binary vectors loaded with engineered CRISPR-Cas12a systems into rice calli and subsequent sequencing revealed that a modified tRNA-crRNA array not only efficiently achieved rice multiplex genome editing, but also successfully edited target sites that were not edited by the crRNA array. This improvement contributes to the application of the CRISPR-Cas12a system in plant genome editing, especially for genomic loci that have hitherto been difficult to edit.

关键词： crRNA CRISPR-Cas12a tRNA-crRNA array Genome editing Editing efficiency

Expanding the Range of CRISPR/Cas9 Genome Editing in Rice

在线全文

学校读者我要写书评

暂无评论

Molecular Plant 2016年第6期9卷 943-945页

作者： xixun hu Chun Wang Yaping FU Qing Liu Xiaozhen Jiao Kejian Wang State Key Laboratory of Rice Biology China National Rice Research Institute Chinese Academy of Agricultural Sciences Hangzhou 310006 China These authors contributed equally to this article.

Dear Editor The CRISPR/Cas9 system has emerged as a versatile molecular tool for genome editing in various organisms, including plants. In this system, the specificity of Cas9-directed DNA cleavage strictly requires the presence of a chimeric single guide RNA （sgRNA） and a short trinucleotide protospacer adjacent motif （PAM） in the genome （Anders et al., 2014; Sternberg et al., 2014）. To date, the Cas9 used in plants has only been shown to recognize PAM sequences in the canonical form NGG （Li et al., 2013; Miao et al., 2013; Shan et al., 2013; Ma et al., 2015b）. As such, the range of sequences for genome editing in plants is limited to sites containing an NGG motif. Many attempts have been made to overcome this constraint, and recent work has revealed that Cas9 can be modified to recognize alternative PAM sequences in zebrafish and human cells （Kleinstiver et al., 2015）. However, the targeting range limitations of the CRISPR/ Cas9 systems in plants have yet to be resolved. The widely cultivated plant rice is not only an important food crop but also a model crop plant because of its relatively small genome and relative ease of transformation. To expand the range of genome editing in rice, we generated two Cas9 variants as reported （Kleinstiver et al., 2015） and investigated their genome editing performance in rice. We demonstrate that Cas9 can be engineered to target sites containing alternative non-canonical PAMs in rice, which significantly broadens the range of genorne editing.

关键词： genome presence trinucleotide

Robust genome editing of CRISPR-Cas9 at NAG PAMs in rice

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Science China(Life Sciences) 2018年第1期61卷 122-125页

作者： Xiangbing Meng xixun hu Qing Liu Xiaoguang Song Caixia Gao Jiayang Li Kejian Wang State Key Laboratory of Plant Genomics and National Center for Plant Gene Research (Beijing) Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing 100101 China State Key Laboratory of Rtee Btology China National Rice Research Institute Chinese Academy of Agricultural Sciences Hangzhou 310006 China State Key Laboratory of Plant Cell and Chromosome Engineering and Center for Genome Editing Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing 100101 China University of Chinese Academy of Sciences Beijing 100049 China

Dear Editor,The CRISPR-Cas9(clustered regularly interspaced short palindromic repeats/Cas9)system has been widely used for a variety of applications,including targeted gene knockout,gene insertion,gene replacement and base editing.Despite its wide use,the genome editing using CRISPR-Cas9 is performed almost exclusively at sites containing canonical NGG protospacer adjacent motifs(PAMs).To overcome the PAM constraint of the CRISPR-Cas9 system,many attempts have been made to develop various Cas9 orthologs

关键词： PAM 染色体编辑米饭柔韧基因插入应用程序系统

Cas9蛋白变体VQR高效识别水稻NGAC前间区序列邻近基序

在线全文

学校读者我要写书评

暂无评论

遗传 2018年第12期40卷 1112-1119页

作者：辛高伟胡熙璕王克剑王兴春山西农业大学生命科学学院太谷030801 中国农业科学院中国水稻研究所水稻生物学国家重点实验室杭州310006

成簇的规律间隔短回文重复序列及CRISPR相关蛋白(clustered regularly interspaced short palindromic repeats/CRISPR-associated9,CRISPR/Cas9)系统是近年来发展起来并被广泛应用的第三代基因组编辑工具。但是,该系统的酿脓链球菌Cas9(Streptococcus pyogenes, SpCas9)仅能识别NGG前间区序列邻近基序(protospacer adjacent motif, PAM),极大地限制了基因组编辑的范围。SpCas9变体VQR(D1135V/R1335Q/T1337R)在水稻中可识别NGAA、NGAG和NGAT PAM,但尚不清楚是否能识别NGAC PAM。本研究利用改进后的CRISPR/VQR系统对水稻中3个相对低效的VQR靶位点NAL1-Q1、NAL1-Q2和LPA1-Q进行了编辑,结果表明改进后的CRISPR/VQR系统可以高效编辑这3个靶位点,编辑效率分别为9.75%、43.90%和29.26%。为了明确改进后的CRISPR/VQR系统对NGAC PAM的识别情况,本研究选择水稻叶片宽度调控基因NARROW LEAF 1 (NAL1)中的NAL-C位点和蜡质合成基因GLOSSY1(GL1)中的GL1-C位点进行基因编辑,并获得57株转基因水稻。靶位点PCR扩增及测序结果表明,NAL1-C和GL1-C靶标位点突变的植株分别为27株和44株,突变率分别为47.36%和77.19%;其中NAL1-C/GL1-C双突变植株为26株,双突变率为45.61%。进一步分析表明,CRISPR/VQR系统造成的突变有4种类型,分别为杂合突变、双等位突变、嵌合体突变和纯合突变,其中以杂合突变和双等位突变为主。这些结果表明,改进的CRISPR/VQR系统可以高效编辑水稻NGAC PAM位点,并产生丰富的突变类型。本研究为水稻及其他植物相关基因NGAC PAM位点的编辑提供了理论依据。

关键词：基因编辑 CRISPR/Cas9 VQR NGAC前间区序列邻近基序水稻

Cross-linking porcine peritoneum by oxidized konjac glucomannan:a novel method to improve the properties of cardiovascular substitute material

在线全文

学校读者我要写书评

暂无评论

Collagen and Leather

Collagen and Leather 2023年第3期5卷 106-122页

作者： Xu Peng Li Li Jiaqi Xing Can Cheng Mengyue hu Yihao Luo Shubin Shi Yan Liu Zhihui Cui xixun Yu College of Polymer Science and Engineering Sichuan UniversityChengdu 610065People's Republic of China Experimental and Research Animal Institute Sichuan UniversityChengdu 610065People's Republic of China Department of Oncology Hematology Western Theater Command Air Force HospitalChengdu 610021People's Republic of China Max Planck Institute for Polymer Research 55128 MainzGermany Department of Growth and Reproduction Copenhagen UniversityCopenhagenDenmark

The use of natural polysaccharide crosslinkers for decellularized matrices is an effective approach to prepare cardiovascular substitute materials.In this research,NalO_(4)was applied to oxidize konjac glucomannan to prepare the polysaccharide crosslinker oxidized konjac glucomannan(OKGM).The as-prepared crosslinker was then used to stabilize collagen-rich decellularized porcine peritoneum(DPP)to construct a cardiovascular substitute material(OKGM-fixed DPP).The results demonstrated that compared with GA-fixed DPP and GNP-fixed DPP,3.75%OKGM[1∶1.5(KGM∶NalO_(4))]-fixed DPP demonstrated suitable mechanical properties,as well as good hemocompatibility,excellent anti-calcification capability,and anti-enzymolysis in vitro.Furthermore,3.75%OKGM[1∶1.5(KGM∶NalO_(4))]-fixed DPP was suitable for vascular endothelial cell adhesion and rapid proliferation,and a single layer of endothelial cells was formed on the fifth day of culture.The in vivo experimental results also showed excellent histocompatibility.The current results demonstrted that OKGM was a novel polysaccharide cross-linking reagent for crosslinking natural tissues featured with rich collagen content,and 3.75%OKGM[1∶1.5(KGM∶NalO_(4))]-fixed DPP was a potential cardiovascular substitute material.

关键词： Sodium periodate Oxidized KGM Peritoneum Collagen Cardiovascular substitute material

Comparison of lymph node number and prognosis in gastric cancer patients with perigastric lymph nodes retrieved by surgeons and pathologists

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Chinese Journal of Cancer Research 2016年第5期28卷 511-518页

作者： Lixin Jiang Zengwu Yao Yifei Zhang Jinchen hu Dawei Zhao huiyuan Zhai xixun Wang Zhenbin Zhang Dong Wang Gastrointestinal Surgery Ward I Yantai Yuhuangding Hospital the Affiliated Hospital of Qingdao University Yantai 264000 China

Objective： To compare the numbers of positive and total lymph nodes and prognosis in gastric cancer patients whose perigastric lymph node retrieval was performed by surgeons and pathologists. Methods： We conducted a retrospective analysis of clinical and follow-up data from 1,056 patients who underwent gastric cancer D2 radical lymph node resection between January 2008 and December 2010 in the Gastrointestinal Surgery Department of Yantai Yuhuangding Hospital. The follow-up ended in December 2015. Patients were divided into two groups according to the specialty of physicians who performed the postoperative perigastric lymph node retrieval： the surgeon group （475 cases） and the pathologist group （581 cases）. The numbers of positive and total perigastric lymph nodes and the 3- and 5-year survival were compared between gastric cancer patients in the two groups overall and stratified by TNM stage （AJCC 7th Edition）. Results： Overall, the numbers of positive and total lymph nodes were significantly higher in the surgeon group than in the pathologist group （6.53±4.07 vs. 4.09±3.70, P=0.021; 29.64±11.50 vs. 20.71±8.56, P〈0.001）. Further analysis showed that the total number of lymph nodes in stage Ⅰ patients （19.40±9.62 vs. 15.45±8.59, P=0.011） and the numbers of positive and total lymph nodes in stage Ⅱ（1.38±1.08 vs. 0.87±1.55, P=0.031; 25.35±10.80 vs. 16.75±8.56, P〈0.001） and stage Ⅲ patients （8.11±6.91 vs. 6.66±5.12, P=0.026; 32.34±12.55 vs. 25.45±8.31, P〈0.001） were significantly higher in the surgeon group than in the pathologist group. The survival analysis showed that the 3- and 5-year survival of stage Ⅱ and Ⅲ patients was significantly higher in the surgeon group than in the pathologist group （82.0% vs. 73.1%, 69.5% vs. 61.2%, P=0.038; 49.2% vs. 38.9%, 36.3% vs. 28.0%; P=0.045）. Conclusions： Compared with retrieval performed by pathologists, postoperative perigastrie lymph node retrieval performed by surgeons was associated with significant increase in the total lymph node number of stage Ⅰ patients, the number

关键词： Gastric cancer surgeon and pathologist lymph node retrieval prognosis

神经浸润与胃癌其他临床病理因素的关系及预后分析

在线全文

学校读者我要写书评

暂无评论

中华内分泌外科杂志 2022年第2期16卷 216-220页

作者：刘传绪王熙勋胡金晨姚增武崔伟豪蔡莉姜立新青岛大学附属烟台毓璜顶医院胃肠外一科烟台264000 青岛大学附属烟台毓璜顶医院病理科烟台264000

目的分析神经浸润与胃癌其他临床病理因素的关系及预后的影响。方法回顾性分析2010年1月至2014年12月在青岛大学附属烟台毓璜顶医院行胃癌根治术的665例胃癌患者的临床病理资料,根据患者是否存在神经浸润将其分为神经浸润阳性组和神经... 详细信息

目的分析神经浸润与胃癌其他临床病理因素的关系及预后的影响。方法回顾性分析2010年1月至2014年12月在青岛大学附属烟台毓璜顶医院行胃癌根治术的665例胃癌患者的临床病理资料,根据患者是否存在神经浸润将其分为神经浸润阳性组和神经浸润阴性组。分析神经浸润与胃癌其他临床病理因素的关系及其对胃癌预后的影响,并使用倾向性评分匹配(PSM)消除两组的潜在混淆偏差后,比较两组胃癌患者5年累积生存率的差异。结果胃癌神经浸润发生率为17.0%(113例),二分类Logistic回归分析显示,肿瘤的浸润深度、脉管癌栓是发生胃癌神经浸润的独立影响因素(P均60岁)、肿瘤直径(>4 cm)、Borrmann分型、浸润深度、淋巴结转移、TNM分期、分化程度、脉管癌栓、神经浸润、癌结节、肿瘤部位、切除部位、手术术式均是胃癌患者预后的影响因素(P均60岁)、肿瘤直径(>4 cm)、浸润深度、淋巴结转移、脉管癌栓阳性是影响胃癌患者预后的独立危险因素(P<0.05),但神经浸润在多因素分析中不能成为胃癌不良预后的独立影响因素。在进行倾向性匹配评分之后进行生存分析,发现神经浸润阳性组和神经浸润阴性组术后五年生存率差异无统计学意义(34.6%vs 43.0%)(χ^(2)=1.713,P=0.191),两者的生存曲线分析无明显差异。结论神经浸润的胃癌患者预后大多不良,但神经浸润不能作为胃癌预后的独立预后因素。

关键词：胃癌神经浸润预后 TNM分期

头侧优先逆时针顺序全结肠系膜切除在腹腔镜右半结肠切除术的应用

在线全文

学校读者我要写书评

暂无评论

中华内分泌外科杂志 2022年第6期16卷 645-649页

作者：姚增武王熙勋张翼飞胡金晨鉴谧刘传绪于彬崔洪铭赵扬姜立新烟台毓璜顶医院胃肠外一科烟台264000 山东大学济南250100 烟台毓璜顶医院开发区分院(烟台业达医院)普外科烟台264000

目的分析头侧优先,逆时针全结肠系膜切除在腹腔镜根治性右半结肠切除术中的有效性和安全性。方法回顾性分析2020年1月至2020年12月烟台毓璜顶医院胃肠外一科收治的右半结肠癌患者30例,均采用头侧优先,进而尾侧,最后中间入路,逆时针顺序... 详细信息

目的分析头侧优先,逆时针全结肠系膜切除在腹腔镜根治性右半结肠切除术中的有效性和安全性。方法回顾性分析2020年1月至2020年12月烟台毓璜顶医院胃肠外一科收治的右半结肠癌患者30例,均采用头侧优先,进而尾侧,最后中间入路,逆时针顺序全结肠系膜切除的入路进行腹腔镜根治性右半结肠切除术。记录患者的一般临床病理学资料,围手术期资料如手术时间、术中出血量、Henle干及属支损伤出血例数,是否中转开腹、术后病理学资料(TNM分期,总清扫淋巴结数目及转移性淋巴结数目)、术后恢复(术后排气时间、进食流质、拔除引流管及住院时间)以及并发症(如出血、吻合口瘘、二次手术、淋巴漏、肺部感染、腹腔感染、切口感染等)。术后1年采用电话或门诊随访等方法进行随访。结果总手术时间(197.80±31.20)min,范围150~275min,术中出血量(58.33±30.30)ml,范围10~100 ml。无术中Henle干属支损伤出血及中转开腹病例。术后排气时间(2.97±0.67)d,范围2~4d;术后进食流质时间(3.67±0.76)d,范围3~5 d;术后拔除引流管时间(6.60±4.00)d,范围4~25 d;术后住院时间为(7.87±3.94)d,范围5~26 d。pTNM分期:Ⅰ期9例、ⅡA期5例、ⅡB期1例、ⅢA期6例、ⅢB期4例、ⅢC期5例。清扫总淋巴结数目(29.50±8.18)枚,范围19~51枚;转移淋巴结数目(1.40±1.77)枚,范围0~6枚。术后并发症:切口感染1例,淋巴漏2例,肺部感染1例,吻合口瘘1例,经过保守治疗均治愈出院。随访期间未发现肿瘤复发或转移,无一例患者死亡。结论头侧优先,逆时针顺序全结肠系膜切除在腹腔镜根治性右半结肠切除术安全有效。

关键词：结肠癌头侧优先逆时针全结肠系膜切除术腹腔镜右半结肠切除术