CRISPR-Cas9 是一种基因编辑技术，通过结合 sgRNA 和 Cas9 蛋白来实现特异性基因靶位点的识别和切割。 sgRNA 通过与目标 DNA 序列互补配对，指导 Cas9 蛋白精确切割目标位点。

sgRNA是一个短的合成RNA，只有20bp大小，可以与Cas9蛋白结合，所以在设计sgRNA之前，应在基因组上寻找 PAM序列 （PAM：Protospacer Adjacent Motif），PAM序列是有固定形式的，来自于不同菌种属的Cas，它的PAM序列形式是不一样的。

2023年8月21日 · 单sgRNA敲除系统：在编码外显子上设计1个sgRNA，依赖细胞本身的非同源末端连接(NHEJ)机制引入或缺失碱基，造成移码突变，从最终得到的混合克隆中筛选符合实验要求的敲除阳性克隆。

2024年8月13日 · 我们通常说的gRNA设计，其实设计的是1-20位，也就是Spacer部分，其余的部分是固定的，称为sgRNA scaffold。 不同种属来源的Cas9的sgRNA scaffold序列不一样。 SpCas9（常用的有两个版本）

2022年6月1日 · SgRNA engineering enables more powerful and versatile CRISPR/Cas9 systems. The sgRNA engineering strategies are summarized in a structural manner. Optimization of sgRNA sequences leads to better genome editing performance. Engineering sgRNA with extra modules expands CRISPR-based functional assays.

This review highlights representative studies that optimize the sgRNA nucleotide sequences for improved genome editing performance (e.g. activity and specificity) as well as add extra aptamers and end extensions for expanded CRISPR-based functional assays (e.g. transcriptional regulation, genome imaging, and prime editor).

2021年10月4日 · Prime editing enables the installation of virtually any combination of point mutations, small insertions or small deletions in the DNA of living cells. A prime editing guide RNA (pegRNA) directs...

2025年3月22日 · Introduction. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) gene editing system has been repurposed to realize live-cell genomic imaging, achieved by inactivating the protein's nuclease activity (dCas9) without abolishing single-guide RNA (sgRNA)-mediated DNA binding [].To light up a specific locus, either dCas9 or sgRNA has ...

2025年2月26日 · To assess sgRNA distribution, the sgRNA target region was amplified using primers that append Illumina sequencing adapters on the 5′ and 3′ ends of the amplicon, as well as a random nucleotide stagger and unique demultiplexing barcode on the 5′ end. ... Corces MR, Granja JM, Shams S, Louie BH, Seoane JA, Zhou W, et al. The chromatin ...

3 天之前 · The sgRNA library encompassing 907 different gRNAs (Twist Bioscience) was cloned into a modified CROP-seq-opti vector (Addgene plasmid #106280) with the puromycin selection marker replaced with ...