{{'Search' | translate}}

Tips LTS 1000 µL 768/8 GPS-L1000

提示LTS 1000 μL768/8

Company: Mettler-Toledo International
Catalog#: GPS-L1000
Other protocol()

Method for Multiplexing CRISPR/Cas9 in Saccharomyces cerevisiae Using Artificial Target DNA Sequences
[Abstract]  Genome manipulation has become more accessible given the advent of the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) editing technology. The Cas9 endonuclease binds a single stranded (single guide) RNA (sgRNA) fragment that recruits the complex to a corresponding genomic target sequence where it induces a double stranded break. Eukaryotic repair systems allow for the introduction of exogenous DNA, repair of existing mutations, or deletion of endogenous gene products. Targeting of Cas9 to multiple genomic positions (termed ‘multiplexing’) is achieved by the expression of multiple sgRNAs within the same nucleus. However, an ongoing concern of the CRISPR field has been the accidental targeting of Cas9 to alternative (‘off-target’) DNA locations within a genome. We ... [摘要]  鉴于CRISPR(集群定期间隔短回归重复)编辑技术的出现,基因组操纵变得更加易于使用。 Cas9核酸内切酶将募集复合物的单链(单向导)RNA(sgRNA)片段结合到相应的基因组靶序列,引发双链断裂。真核修复系统允许引入外源DNA,修复现有突变或内源基因产物的缺失。通过在同一核内表达多个sgRNA来实现Cas9对多个基因组位置的定位(称为“多重”)。然而,CRISPR领域的持续关注是将Cas9意外地定位到基因组内的替代(“脱靶”)DNA位置。我们将安装的人造Cas9靶序列的使用(称为人造基因座上的Cas9复制)描述为允许(i)与单个sgRNA复用的酵母基因组中的用途; (ii)减少/消除可能的脱靶效应,以及(iii)精确控制预定目标序列的放置。
【背景】CRISPR(集群定期间隔回归重复)机制已经在原核生物中演变为具有很高精度编辑任何基因组的能力的原始适应性免疫系统(Jinek等,2012; Sorek等,2013)。这种生物技术需要使用来自化脓性链球菌(或othologous物种)的内切核酸酶(Cas9),单个RNA'引导'序列和外源供体DNA(如果需要)。仅在短短几年内,CRISPR / ...

Scratch Wound Healing Assay
[Abstract]  The scratch wound healing assay has been widely adapted and modified to study the effects of a variety of experimental conditions, for instance, gene knockdown or chemical exposure, on mammalian cell migration and proliferation. In a typical scratch wound healing assay, a “wound gap” in a cell monolayer is created by scratching, and the “healing” of this gap by cell migration and growth towards the center of the gap is monitored and often quantitated. Factors that alter the motility and/or growth of the cells can lead to increased or decreased rate of “healing” of the gap (Lampugnani, 1999). This assay is simple, inexpensive, and experimental conditions can be easily adjusted for different purposes. The assay can also be used for a high-throughput screen platform if an automated system ... [摘要]  划伤伤口愈合测定法已经广泛适应和修改以研究多种实验条件例如基因敲低或化学暴露对哺乳动物细胞迁移和增殖的影响。 在典型的伤口创伤愈合测定中,通过刮擦产生细胞单层中的"伤口间隙",并且监测并常常定量通过细胞迁移和朝向间隙中心的生长的该间隙的"愈合"。 改变细胞运动性和/或生长的因素可导致间隙"愈合"的增加或减少的速率(Lampugnani,1999)。 该测定简单,便宜,并且可以容易地针对不同目的调整实验条件。 如果使用自动化系统,该测定也可以用于高通量筛选平台(Yarrow和Perlman,2004)。