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| 1 | Hi-TOM: a platform for high-throughput tracking of mutations induced by CRISPR/Cas systems显示文摘The CRISPR/Cas system has been extensively applied to make precise genetic modifications in various organisms. Despite its importance and widespread use, large-scale mutation screening remains time-consuming, labour-intensive and costly. Here, we developed Hi-TOM(available at http://gffzz103f3ce5d9b14419hk5ovuq0wfuuu6bwx.ffgz.tsg.suse.edu.cn/hi-tom/), an online tool to track the mutations with precise percentage for multiple samples and multiple target sites. We also described a corresponding next-generation sequencing(NGS) library construction strategy by fixing the bridge sequences and barcoding primers. Analysis of the samples from rice, hexaploid wheat and human cells reveals that the Hi-TOM tool has high reliability and sensitivity in tracking various mutations, especially complex chimeric mutations frequently induced by genome editing. Hi-TOM does not require special design of barcode primers,cumbersome parameter configuration or additional data analysis. Thus, the streamlined NGS library construction and comprehensive result output make Hi-TOM particularly suitable for high-throughput identification of all types of mutations induced by CRISPR/Cas systems. | Qing Liu Chun Wang Xiaozhen Jiao Huawei Zhang Lili Song Yanxin Li Caixia Gao Kejian Wang | 2019 | Science China(Life Sciences)2019，62，1： | 37 |
| 2 | Genome-editing technologies: the gap between application and policy显示文摘Genome editing is a new technology for manipulating genomic DNA sequences at specific sites,and researchers worldwide are rapidly developing new variants of genomeediting applications that target mRNAs,viruses,chromatin structure,and other cellular processes. | Gousi Li Yao-Guang Liu Yuanling Chen | 2019 | Science China(Life Sciences)2019，62，11： | 9 |
| 3 | The evolving CRISPR technology显示文摘Adva nces in genome editi ng tech no logy have revoluti on ized basic and applied biology research in recent years,particu?larly due to the newly emerged CRISPR/Cas technique(Ren et al.,2017b).The classical CRISPR/Cas system was derived from a bacterial defense system,which consists of a single guide RNA(sgRNA)for precise targeting and one Cas protein for DNA binding and nuclease activity.Relying on high precision of CRISPR/Cas system,multiple functions have been exploited in addition to original nuclease activity,such as genome base editing,gene knock-down and activation,chromatin imaging systems etc. | Meng Yan Jinsong Li | 2019 | Protein & Cell2019，10，11： | 3 |
| 4 | A new and promising application of gene editing: CRISPR-controlled smart materials for tissue engineering,bioelectronics, and diagnostics显示文摘A recent study published in the journal Science by Collins et al.proposed a programmable clustered regularly interspaced short palindromic repeats(CRISPR)-responsive smart material containing the CRISPR-associated nuclease,Casl2a,and hydrogels containing DNA to deliver biological information via changes in material properties(English et al.,2019). | Weiqi Hong Mengyuan Huang Yuquan Wei Xiawei Wei | 2019 | Science China(Life Sciences)2019，62，11： | 3 |
| 5 | C1orf106, an innate immunity activator, is amplified in breast cancer and is required for basal-like/luminal progenitor fate decision显示文摘Basal-like breast cancer with a luminal progenitor gene expression profile is an aggressive subtype of breast cancer with a poorer prognosis compared with other subtypes.However,genes that specifically promote basal-like breast cancer development remain largely unknown.Here,we report that a novel gene C1orf106 plays an important role in maintaining the feature of basal-like/luminal progenitors.C1orf106 is frequently amplified and overexpressed in basal-like breast cancer and is associated with a poor outcome in patients.In human TCGA database,C1orf106 expression was correlated with upregulation of ELF5 and downregulation of GATA3,two transcription factors that regulate mammary gland stem cell fate.Enhanced expression of C1orf106 promotes tumor progression and expression of basal-like/luminal progenitor marker ELF5;depletion of C1orf106 suppresses tumorigenesis and expression of basal-like/luminal progenitor marker GATA3.These findings suggest that C1orf106 maintains the basal-like/luminal progenitor character through balancing the expression of ELF5 and GATA3.Taken together,we demonstrated that C1orf106 is an important regulator for basal-like/luminal progenitors and targeting C1orf106 is of therapeutic value for breast cancer. | Ji Ma Cheng Liu Decao Yang Jiagui Song Jing Zhang Tianzhuo Wang Mengyuan Wang Weizhi Xu Xueying Li Shigang Ding Jun Zhan Hongquan Zhang | 2019 | Science China(Life Sciences)2019，62，9： | 2 |
| 6 | Multiplex genome editing using a dCas9-cytidine deaminase fusion in Streptomyces显示文摘CRISPR/Cas-mediated genome editing has greatly facilitated the study of gene function in Streptomyces. However, it could not be efficiently employed in streptomycetes with low homologous recombination(HR) ability. Here, a deaminase-assisted base editor d Cas9-CDA-ULstr was developed in Streptomyces, which comprises the nuclease-deficient Cas9(dCas9), the cytidine deaminase from Petromyzon marinus(PmCDA1), the uracil DNA glycosylase inhibitor(UGI) and the protein degradation tag(LVA tag). Using d Cas9-CDA-ULstr , we achieved single-, double-and triple-point mutations(cytosine-to-thymine substitutions)at target sites in Streptomyces coelicolor with efficiency up to 100%, 60% and 20%, respectively. This base editor was also demonstrated to be highly efficient for base editing in the industrial strain, Streptomyces rapamycinicus, which produces the immunosuppressive agent rapamycin. Compared with base editors derived from the cytidine deaminase rAPOBEC1, the PmCDA1-assisted base editor dCas9-CDA-ULstr could edit cytosines preceded by guanosines with high efficiency, which is a great advantage for editing Streptomyces genomes(with high GC content). Collectively, the base editor dCas9-CDA-ULstr could be employed for efficient multiplex genome editing in Streptomyces. Since the d Cas9-CDA-ULstr -based genome editing is independent of HR-mediated DNA repair, we believe this technology will greatly facilitate functional genome research and metabolic engineering in Streptomyces strains with weak HR ability. | Yawei Zhao Jinzhong Tian Guosong Zheng Jun Chen Chuanwen Sun Zhongyi Yang Andrei A.Zimin Weihong Jiang Zixin Deng Zhijun Wang Yinhua Lu | 2020 | Science China(Life Sciences)2020，63，7： | 2 |
| 7 | Anything impossible with CRISPR/Cas9?显示文摘Since the adaptation of the bacterial CRISPR/Cas9 system to eukaryotic cells,this'magic'tool has now been dramatically modified and applied to all kinds of biomedical research,clinics and agriculture(Jiao and Gao,2016).The system was first reported to rapidly,specifically and effectively target genomic sequences in human cells(Cong et al.,2013;Jinek et al.,2013).Since then it attracted scientists’ | Renjie Jiao Caixia Gao | 2017 | Science China(Life Sciences)2017，60，5： | 1 |
| 8 | 基于“人造精子细胞”的基因组标签计划显示文摘蛋白质是一切生命活动的基础,蛋白质研究特别是生命活动中蛋白质功能网络调控的揭示成为继人类基因组计划后广受关注的重要科学问题.然而,大规模开展蛋白质研究缺乏一个高效、标准的平台,构建基因组范围的蛋白质标签模式生物资源平台是解决这一问题的关键.虽然在酵母、线虫、果蝇等模式生物上已经实现了基因组范围的蛋白质标签文库的构建,但是在小鼠个体水平上,基因组范围的蛋白质标签的构建仍然困难重重.基因组标签计划(Genome tagging project, GTP)的提出是基于孤雄单倍体胚胎干细胞(又称为'人造精子细胞')介导的半克隆技术和CRISPR-Cas9基因编辑技术的发展:首先在'人造精子细胞'上利用CRISPR-Cas9技术对特定编码蛋白质基因进行标签序列的原位插入获得相应的细胞系,进一步通过卵子注射即能获得携带标签的小鼠模型.因此,该策略简单、高效、成本低,可以用于基因组范围制备蛋白质标签小鼠. GTP的目标是给小鼠中的每个蛋白质带上标签, GTP的实施有望实现以一套标准化的方法来对每个蛋白质进行在体研究,将促进生命科学的发展. | 常柏然 李劲松 | 2020 | 中国科学:生命科学2020，50，5： | 1 |
| 9 | Reduction of BCL11A in hematopoietic stem cells through gene editing:new strategy to ameliorate the severe b-globin disorders sickle cell disease显示文摘Site-specific gene editing is of great importance in precise medicine. Two conventional genome editing methods, Zine finger nucleases (ZFNs) and transcription activator-like effector nucleases(TALENs), are based on protein-DNA recognition, with tedious work in constructing target protein [1,2]. Developed from immune response of bacteria, CRISPR/Cas9 has been widely investigated as a promising tool for therapeutic genome editing in clinical settings nowadays[3,4]. | Weiqi Hong Mengyuan Huang Yuquan Wei Xiawei Wei | 2019 | Science Bulletin2019，64，21： | 0 |
| 10 | Ⅰ型神经纤维瘤病的基因治疗策略与前景显示文摘目的总结Ⅰ型神经纤维瘤病(neurofibromatosis type 1,NF1)基因治疗策略和相关研究进展。方法查阅近年来国内外关于NF1基因治疗的文献,对NF1基因结构、功能及其突变进行分析,并从转基因治疗和基因编辑两方面对基因治疗策略进行深入总结。结果NF1是由抑癌基因NF1基因突变引起的常染色体显性遗传肿瘤性疾病,因神经纤维瘤蛋白功能受损而致病,临床表现复杂,目前尚无法根治。NF1的基因治疗策略仍处于研究和开发阶段。现有NF1转基因疗法主要是在神经纤维瘤蛋白缺陷的细胞中构建并表达Ras-GTP酶激活蛋白相关结构域基因片段,证实了NF1基因治疗的可行性;未来研究方向主要聚焦于使用拆分载体递送系统,增加单一载体的运载量,以及开发新的递送系统用于靶向递送全长NF1 cDNA。此外,新一代基因编辑工具在NF1等单基因遗传病的治疗领域应用潜力巨大,但效率和安全性尚需进一步验证。结论基因治疗,包括转基因和基因编辑两大技术,有望成为NF1患者重要的新型治疗手段。 | 郑婷婷 朱倍瑶 王智超 李青峰 | 2024 | 中国修复重建外科杂志2024，38，1： | 0 |
| 11 | CRISPR/Cas9系统介导的斑马鱼kctd1纯合突变体的构建显示文摘KCTD1 (potassium channel tetramerization domain containing 1)属于钾离子通道四聚化结构域蛋白基因家族的成员,该家族成员的N端具有保守的BTB(Bric-a-brack, Tram-track, Broad complex)结构域和一个钾离子四聚体通道结构域(K-ketra),但C端是多变的。人类 KCTD 基因家族成员的突变和表达异常与多种疾病相关。研究选取人类皮肤-耳朵-乳头综合征(Scalp-Ear-Nipple Syndrome,SEN)致病基因 KCTD1 在斑马鱼( Danio rerio )中的直系同源基因 kctd1 ,利用CRISPR/Cas9技术进行基因敲除,通过F 1 剪尾检测和F 1 内交,成功获得 kctd1 F 2 纯合突变体。获得的突变体类型为+7 bp和-8 bp。研究结果对于人类 KCTD1 基因功能以及KCTD1致病机理的研究具有参考价值。 | 刘姣 黄姣 胡金萌 李伟明 任建峰 | 2019 | 生物学杂志2019，36，5： | 0 |
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