分子生物学

分类

    现刊
    GeneWeld: Efficient Targeted Integration Directed by Short Homology in Zebrafish
    GeneWeld:斑马鱼短同源性诱导的高效靶向整合
    [Abstract]

    Efficient precision genome engineering requires high frequency and specificity of integration at the genomic target site. Multiple design strategies for zebrafish gene targeting have previously been reported with widely varying frequencies for germline recovery of integration alleles. The GeneWeld protocol and pGTag (plasmids for Gene Tagging)

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    Defined Mutant Library Sequencing (DML-Seq) for Identification of Conditional Essential Genes
    确定突变体文库测序(DML-Seq)用于鉴定的条件必需基因
    作者:Shuai Shao, Lifan Wei, Feng Xia, Yuanxing Zhang and Qiyao Wang日期:03/05/2021,浏览量:2079,Q&A: 0
    [Abstract]

    Transposon insertion sequencing (TIS) is an emerging technique which utilizes a massive transposon mutant library to screen specific phenotype and determine the conditional essential genetic requirements for bacterial fitness under distinct conditions combined with high-throughput parallel sequencing technology. Compared with a massive mutant

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    Rapid Genome Engineering of Pseudomonas Assisted by Fluorescent Markers and Tractable Curing of Plasmids
    受荧光标记和易于固化质粒影响的假单胞菌快速基因组工程
    作者:Daniel C. Volke, Nicolas T. Wirth and Pablo I. Nikel日期:02/20/2021,浏览量:2834,Q&A: 0
    [Abstract]

    Precise genome engineering has become a commonplace technique for metabolic engineering. Also, insertion, deletion and alteration of genes and other functional DNA sequences are essential for understanding and engineering cells. Several techniques have been developed to this end (e.g., CRISPR/Cas-assisted methods, homologous recombination, or λ

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    Screening Method for CRISPR/Cas9 Inhibition of a Human DNA Virus: Herpes Simplex Virus
    人单纯疱疹病毒CRISPR/Cas9抑制作用的筛选方法
    [Abstract] The efficiency of cleavage of individual CRISPR/Cas9-sgRNAs remains difficult to predict based on the CRISPR target sequence alone. Different intracellular environments (dependent on cell type or cell cycle state for example) may affect sgRNA efficiency by altering accessibility of genomic DNA through DNA modifications such as epigenetic marks and ...
    Construction of Antisense RNA-mediated Gene Knock-down Strains in the Cyanobacterium Anabaena sp. PCC 7120
    反义RNA介导的鱼腥藻PCC 7120基因敲除株的构建
    作者:Amit Srivastava, Anand Ballal, Karl Forchhammer and Anil Kumar Tripathi日期:02/20/2020,浏览量:2978,Q&A: 0
    [Abstract] Anabaena sp. PCC 7120 (hereafter Anabaena) is a model cyanobacterium to study nitrogen fixation, cellular differentiation and several other key biological functions that are analogous in plants. As with any other organism, many genes in Anabaena encode an essential life function and hence cannot be deleted, causing a ...
    High-throughput Site-directed Scanning Mutagenesis Using a Two-fragment PCR Approach
    利用双片段PCR法进行高通量定点扫描突变
    作者:Franziska M. Heydenreich, Tamara Miljuš, Dalibor Milić and Dmitry B. Veprintsev日期:01/05/2020,浏览量:3358,Q&A: 0
    [Abstract] Site-directed scanning mutagenesis is a useful tool applied in studying protein function and designing proteins with new properties, such as increased stability or enzymatic activity. Creating a systematic library of hundreds of site-directed mutants is still a demanding and expensive task. The established protocols for making such libraries ...
    In vitro Generation of CRISPR-Cas9 Complexes with Covalently Bound Repair Templates for Genome Editing in Mammalian Cells
    利用共价修复模板体外制备用于哺乳动物细胞中基因组编辑的CRISPR-Cas9复合物
    [Abstract] The CRISPR-Cas9 system is a powerful genome-editing tool that promises application for gene editing therapies. The Cas9 nuclease is directed to the DNA by a programmable single guide (sg)RNA, and introduces a site-specific double-stranded break (DSB). In mammalian cells, DSBs are either repaired by non-homologous end joining (NHEJ), generating ...
    Generation of Gene Knockout and Gene Replacement with Complete Removal of Full-length Endogenous Transcript Using CRISPR-Trap
    通过CRISPR-Trap完全移除内源全长转录本进行基因敲除和基因置换
    作者:Jonas Mechtersheimer, Stefan Reber and Marc-David Ruepp日期:10/20/2018,浏览量:5851,Q&A: 0
    [Abstract] This protocol describes the application of the CRISPR-Trap from designing of the gene targeting strategy to validation of successfully edited clones that was validated on various human cell lines, among them human induced pluripotent stem cells (hiPSCs). The advantage of CRISPR-Trap over conventional approaches is the complete removal of any ...
    Delivery of the Cas9 or TevCas9 System into Phaeodactylum tricornutum via Conjugation of Plasmids from a Bacterial Donor
    通过细菌供体质粒接合转移的方法将Cas9或 TevCas9系统导入三角褐指藻
    作者:Helen Wang, Samuel S. Slattery, Bogumil J. Karas and David R. Edgell日期:08/20/2018,浏览量:5579,Q&A: 2
    [Abstract] Diatoms are an ecologically important group of eukaryotic microalgae with properties that make them attractive for biotechnological applications such as biofuels, foods, cosmetics and pharmaceuticals. Phaeodactylum tricornutum is a model diatom with defined culture conditions, but routine genetic manipulations are hindered by a lack of ...
    Random Insertional Mutagenesis of a Serotype 2 Dengue Virus Clone
    2型登革病毒克隆的随机插入诱变
    作者:Jeffrey W. Perry and Andrew W. Tai日期:08/20/2018,浏览量:3832,Q&A: 0
    [Abstract] Protein tagging is a powerful method of investigating protein function. However, modifying positive-strand RNA virus proteins in the context of viral infection can be particularly difficult as their compact genomes and multifunctional proteins mean even small changes can inactivate or attenuate the virus. Although targeted approaches to ...