Cas13d的guide RNA设计可以参考这个网站Cas13design (nygenome.org),首页有一些参考文献可以读一下。
Cas13d酶的一个有趣的特点就是它们的切割模式:当guide结合在目标RNA上时,EsCas13d产生非常相似的切割产物,这表明该酶在离目标区域可预测的距离上没有发生切割。Konermann等人的研究表明EsCas13d倾向于在尿嘧啶处切割,但对这种切割模式的更详细的探索是十分必要的。 Konermann等比较了CasRx与多种RNA调控方法:小发...
他们将Cas13d系统和靶向Pten基因的guide RNA导入到小鼠肝脏细胞中,成功在肝脏细胞中沉默了Pten基因,并通过增强下游蛋白AKT的磷酸化,影响了相关糖脂代谢相关基因的表达。这说明Cas13d可以应用于模拟体内代谢信号的调控。此外,研究者利用Cas13d系统,成功地降低了小鼠血液中的胆固醇水平。低密度脂蛋白胆固醇(low-density li...
Wessels, HH. et al. (2020) Massively parallel Cas13 screens reveal principles for guide RNA design. Nat Biotechnol. 38;722–727. Guo X. et al. (2021) Transcriptome-wide Cas13 guide RNA design for model organisms and viral RNA pathogens. Cell Genomics...
美国科学家团队开发出适应于CRISPR/Cas13系统的guide RNA设计体系,研究成果于2020年3月16日在《自然·生物技术》上在线发表。研究表明,gRNA存在保证其靶向活性的核心区域,其中心位于相对于gRNA 5’端第18个核苷酸,并发现当错配位于靶标位点...
The appropriate statistical tests were chosen on the basis of experimental design after consulting the GraphPad Statistics Guide. Homology analysis Protein sequence for RfxCas13d (accession number: SFX39573.1) was aligned with one of the top hits found using NCBI Protein Blast, Cas13d from a ...
CRISPR–Cas13 systems have recently been used for targeted RNA degradation in various organisms. However, collateral degradation of bystander RNAs has limited their in vivo applications. Here, we design a dual-fluorescence reporter system for detecting collateral effects and screening Cas13 variants in...
Cas13是一种CRISPR-Cas蛋白,它可以通过形成复合物形式帮助你研究和调控哺乳动物细胞中的RNA。Cas13如果想要发挥功能,它需要搜寻您需要修饰或者靶向的RNA。那么就利用一个较短且具有柔性的RNA就比较容易精准定位到你需要靶向的RNA。这种能够通过碱基互补配对方式将Cas13导向目标分子的RNA被称为引导RNA(guide RNA,gRNA)。
To understand the molecular basis for Cas13d function and explain its compact molecular architecture, we resolved cryoelectron microscopy structures of Cas13d-guide RNA binary complex and Cas13d-guide-target RNA ternary complex to 3.4 and 3.3 Å resolution, respectively. Furthermore, a 6.5 Å...
. In plants, CRISPR/Cas9 efficiently confers viral resistance to host plants. We and others have shown that CRISPR/Cas9 mediates strong interference against various DNA viruses. We have also shown that a single guide RNA targeting a conserved intergenic region in multiple Gemini viruses guides Cas9...