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。
2c). Although we observed translational repression of Rluc with all the gRNAs tested, central placement of the anti-start codon (e.g., gRNA AUG9 and gRNA AUG17) in the spacer had a positive effect on translational repression (Fig. 2d and Supplementary Fig. 2b). A similar design ...
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...
Functional overview of CRISPR/Cas9 and CRISPR/Cas13 systems.aSchematic of Cas9 mechanism in genome editing. This system requires the recruitment of CRISPR-associated protein Cas9 (blue) to the target site recognized by the guide RNA (gRNA: orange). Target site cleavage by Cas9 is ensured by th...
A major challenge in the application of the CRISPR-Cas13d system is to accurately predict its guide-dependent on-target and off-target effect. Here, we perform CRISPR-Cas13d proliferation screens and design a deep learning model, named DeepCas13, to pred
design, each cell stably expresses a specific gRNA that upon induction of Cas13 also integrated into the genome can induce knockdown of the target transcript. Thus, each cell is barcoded by a unique gRNA sequence. The effect of the knockdown on cellular viability could then be judged by ...