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1.
Targeted editing of intronic-splicing silencer enhancement of SMN2 Exon 7 inclusion by CRISPR/Case 9
LIUCHENG WU YI WANG LILI DU GUIQING JI RUI ZHOU ZEYI ZHAO JUN CHEN SHUNXING ZHU 《Biocell》2021,45(6):1501-1507
Spinal muscular atrophy (SMA) is an autosomal recessive hereditary neuromuscular disease. Exon 7 and 8 of survival of motor neuron 1 (SMN1) gene or only exon 7 homology deletion leads to the failure to produce a full-length SMN gene. The copy number of SMN2 gene with high homology of SMN1 affects the degree of disease and was the target gene for targeting therapy, in which splicing silencer in intron 7 was the key to suppress the inclusion of exon 7. In this study, we projected to use CRISPR/Case 9 for the targeted editing of intronic-splicing silencer (ISS) sequence to promote the inclusion of SMN2 exon 7 and increase the production of SMN2 full-length (FL) gene expression. It happens that there was a protospacer adjacent motif (PAM) at one end of the ISS sequence according to the design of sgRNA. The recombinant vector of sgRNA HSMN2 CRISPR/Case 9 was constructed and transfected into HEK293 cells. Sequencing results showed that the ISS sequence could be edited accurately and targeting in the predicted direction, in which deleting small fragments, inserting small amounts and mutation. Quantitative analysis of RT-PCR products by restriction enzyme of DdeI digestion showed that the FL of SMN2 increased by 8% (P < 0.05). In the primary cultured chondrocytes of SMA mice, in which sgRNA HSMN2 CRISPR/Case9 recombinant vector transfection could increase the SMN2 FL gene by 23% (P < 0.05) and significantly improve SMN protein levels (P < 0.05). CRISPR/Case 9 is an effective tool for gene editing and therapy of hereditary diseases, but it is rarely reported in the treatment of SMA diseases. This study shows that CRISPR/Case 9 was first used for the precision target of ISS sequence editing, which can effectively promote the production of SMN2 FL gene expressions, in which there was an important clinical reference value. 相似文献
2.
Marc De Doncker Chloé De Graeve Dr. Jorick Franceus Dr. Koen Beerens Prof. Vladimír Křen Dr. Helena Pelantová Dr. Ronny Vercauteren Prof. Dr. Tom Desmet 《Chembiochem : a European journal of chemical biology》2021,22(23):3319-3325
The substantial increase in DNA sequencing efforts has led to a rapid expansion of available sequences in glycoside hydrolase families. The ever-increasing sequence space presents considerable opportunities for the search for enzymes with novel functionalities. In this work, the sequence-function space of glycoside hydrolase family 94 (GH94) was explored in detail, using a combined approach of phylogenetic analysis and sequence similarity networks. The identification and experimental screening of unknown clusters led to the discovery of an enzyme from the soil bacterium Paenibacillus polymyxa that acts as a 4-O-β-d -glucosyl-d -galactose phosphorylase (GGalP), a specificity that has not been reported to date. Detailed characterization of GGalP revealed that its kinetic parameters were consistent with those of other known phosphorylases. Furthermore, the enzyme could be used for production of the rare disaccharides 4-O-β-d -glucosyl-d -galactose and 4-O-β-d -glucosyl-l -arabinose. Our current work highlights the power of rational sequence space exploration in the search for novel enzyme specificities, as well as the potential of phosphorylases for rare disaccharide synthesis. 相似文献
3.
《全球能源互联网(英文)》2022,5(6):618-626
A novel image sequence-based risk behavior detection method to achieve high-precision risk behavior detection for power maintenance personnel is proposed in this paper. In this method, the original image sequence data is first separated from the foreground and background. Then, the free anchor frame detection method is used in the foreground image to detect the personnel and correct their direction. Finally, human posture nodes are extracted from each frame of the image sequence, which are then used to identify the abnormal behavior of the human. Simulation experiment results demonstrate that the proposed algorithm has significant advantages in terms of the accuracy of human posture node detection and risk behavior identification. 相似文献
4.
Accurate and timely network traffic measurement is essential for network status monitoring, network fault analysis, network intrusion detection, and network security management. With the rapid development of the network, massive network traffic brings severe challenges to network traffic measurement. However, existing measurement methods suffer from many limitations for effectively recording and accurately analyzing big-volume traffic. Recently, sketches, a family of probabilistic data structures that employ hashing technology for summarizing traffic data, have been widely used to solve these problems. However, current literature still lacks a thorough review on sketch-based traffic measurement methods to offer a comprehensive insight on how to apply sketches for fulfilling various traffic measurement tasks. In this paper, we provide a detailed and comprehensive review on the applications of sketches in network traffic measurement. To this end, we classify the network traffic measurement tasks into four categories based on the target of traffic measurement, namely cardinality estimation, flow size estimation, change anomaly detection, and persistent spreader identification. First, we briefly introduce these four types of traffic measurement tasks and discuss the advantages of applying sketches. Then, we propose a series of requirements with regard to the applications of sketches in network traffic measurement. After that, we perform a fine-grained classification for each sketch-based measurement category according to the technologies applied on sketches. During the review, we evaluate the performance, advantages and disadvantages of current sketch-based traffic measurement methods based on the proposed requirements. Through the thorough review, we gain a number of valuable implications that can guide us to choose and design proper traffic measurement methods based on sketches. We also review a number of general sketches that are highly expected in modern network systems to simultaneously perform multiple traffic measurement tasks and discuss their performance based on the proposed requirements. Finally, through our serious review, we summarize a number of open issues and identify several promising research directions. 相似文献
5.
Facial Expression Recognition (FER) is an important subject of human–computer interaction and has long been a research area of great interest. Accurate Facial Expression Sequence Interception (FESI) and discriminative expression feature extraction are two enormous challenges for the video-based FER. This paper proposes a framework of FER for the intercepted video sequences by using feature point movement trend and feature block texture variation. Firstly, the feature points are marked by Active Appearance Model (AAM) and the most representative 24 of them are selected. Secondly, facial expression sequence is intercepted from the face video by determining two key frames whose emotional intensities are minimum and maximum, respectively. Thirdly, the trend curve which represents the Euclidean distance variations between any two selected feature points is fitted, and the slopes of specific points on the trend curve are calculated. Finally, combining Slope Set which is composed by the calculated slopes with the proposed Feature Block Texture Difference (FBTD) which refers to the texture variation of facial patch, the final expressional feature are formed and inputted to One-dimensional Convolution Neural Network (1DCNN) for FER. Five experiments are conducted in this research, and three average FER rates 95.2%, 96.5%, and 97% for Beihang University (BHU) facial expression database, MMI facial expression database, and the combination of two databases, respectively, have shown the significant advantages of the proposed method over the existing ones. 相似文献
6.
吐哈盆地葡北地区“两宽一高”地震勘探技术及应用效果 总被引:1,自引:0,他引:1
吐哈盆地台北凹陷圈闭落实的准确度和储层分布预测的精度一直是制约该区油气发现的关键因素。通过可控震源“两宽一高”地震采集、OVT域时间偏移、地质导向深度偏移初始模型建立及各向异性深度偏移迭代处理,在地震资料的信噪比、频带宽度和分辨率等方面明显提高的基础上,以高精度层序地层学为指导,基于高分辨率地震资料与钻井、测井资料的综合分析,识别出侏罗系6个三级层序,在三级层序格架内将研究区七克台组底部和三间房组顶部砂体识别为两个连续上升半旋回四级层序;通过构造几何属性分析及精细构造分析,揭示了本区“四带、两凹”的构造格局及形成机制;以宽频带多种反演技术为手段,对含油气层段的七克台组和三间房组砂体分布进行了预测;在综合构造分析、储层分析和油藏成藏规律分析的基础上,提出本区油气沿构造带脊线聚集,断裂系统是油气运移的优势路径,七克台组、三间房组储层在葡北构造带发育,向玉果构造带和胜北次凹减薄尖灭,葡北构造带与玉果构造带结合部位的,具有鼻状构造背景的岩性圈闭是获得油气突破的有利地区。 相似文献
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为了有效降低因驾驶员紧急换道行为而诱发的交通事故,提高道路交通事故链阻断效率,提出一种基于高斯混合隐马尔科夫模型(GMM-HMM)和人工神经网络(ANN)的紧急换道行为预测方法。首先利用GMM-HMM对车辆行驶状态以及驾驶行为连续观察序列进行换道意图辨识,采用ANN预测下一时段的驾驶行为,再预测换道过程中的横向加速度变化率,从而判断紧急换道的危险程度。驾驶员在环仿真实验及实车实验结果表明,该方法预测避险成功率达92.83%,实验避险成功率达90.32%。该方法能有效地对紧急换道行为进行提前警告与干预。 相似文献
10.
《中国有色金属学会会刊》2020,30(1):76-89
The distribution of temperature and then the distribution of residual stress and distortion in the stiffened aluminum alloy Al6061-T6 plates under the metal inert gas (MIG) welding process were investigated by three dimensional thermo-mechanical coupled finite element model using Ansys software. The properties of materials were considered temperature-dependent and the filler metal was added to the workpiece by the element birth and death technique. In three modes of current, two different speeds and two various sequences, the distribution of residual stress and distortion were calculated and analyzed. The results showed that increase in welding speed decreased the vertical deflection in the plate, transverse shrinkage and angular distortion of plate and the lateral deflection of stiffener, but increased the maximum longitudinal tensile stress in the plate and stiffener. Furthermore, increase in current increased the residual stress and deformation in the plate and stiffener, and the change in the welding sequence changed the distribution of the distortion in the plate and the stiffener without significant change in the distribution of the longitudinal residual stress. 相似文献