2019年5月7日 · We first establish a viable strategy for augmenting Cas12a guide RNAs (gRNAs) at their 5′ end and then use such 5′ extensions to construct strand displacement gRNAs (SD gRNAs) that can be activated...

2015年8月17日 · Here we report a simple, inexpensive, one-step method that allows for the rapid and efficient cloning of gRNA pairs into expression vectors. We show that this method can be used to generate...

The outlined protocol describes streamlined methods for the efficient and cost-effective generation of Cas9-associated guide RNAs. Two alternative strategies for guide RNA (gRNA) cloning are outlined based on the usage of the Type IIS restriction ...

CRISPR/Cas9是第二类中依赖gRNA引导单一具有内切核酸酶活心的 cas9蛋白 形成复合物（sgRNA的3’端是具有结合cas9蛋白的双链RNA结构，负责招募cas9到靶向区域），实现对核酸DNA效应，依赖细胞中自身的 同源重组 （HDR）或者 非同源末端连接 （NHEJ）的途径，实现目 …

2020年1月3日 · Here, we introduce post-synthetic masking and chemical activation of guide RNA (gRNA) to controlling CRISPR systems. An RNA structure profiling probe (2-azidomethylnicotinic acid imidazolide) is...

CRISPR protocol (gRNA cloning) by Huang Last updated: 08/21/2014 . Laboratory of Cancer Genetics and Epigenetics, Cancer Institute of Fudan University Shanghai Cancer Center -5-

The ease of gRNA design and synthesis, as well as the ability to target almost any genomic locus, makes CRISPR an ideal genome engineering system for large-scale, forward genetic screening. CRISPR can make highly specific, permanent genetic modifications to …

2024年4月25日 · This protocol outlines the construction of a plant transformation plasmid to express both the Cas9 nuclease and individual guide RNA (gRNA), facilitating the induction of double-stranded breaks (DSBs) in DNA and subsequent imprecise repair …

Here, we present a workflow to perform specific, efficient, and cost-effective mRNA knockdown in zebrafish embryos using our in vivo opt... ... Check the gRNA integrity by running an aliquot in a...

Under the inspiration of Jacob L. Litke and Samie R. Jaffrey, we propose a novel gRNA design that could achieve rapid circular gRNA assembly inside the cells. This circular design consists of the gRNA of interested flanked by Twister ribozymes.