2017年2月16日 · We establish that these CRISPR-Cas systems can achieve RNA interference when heterologously expressed. Through a combination of biochemical and genetic experiments, we show that Cas13b processes its own CRISPR array with short and long direct repeats, cleaves target RNA, and exhibits collateral RNase activity.

Broad研究所张锋团队基于dLwaCas13a-msfGFP融合蛋白，借助核定位序列、锌指蛋白及KRAB结构域，利用负反馈调节机制提升信噪比。 该系统信噪比有限，因此只被应用于表达丰度高的 ACTB mRNA [5] （图3.2）。 针对lncRNA与亚细胞结构的研究，中科院陈玲玲团队开发lncRNA示踪技术。 该团队通过对不同dCas13蛋白进行系统筛选，发现dPspCas13b与dPguCas13b蛋白靶向性最好。 随后，团队对crRNA长度、位点、荧光蛋白融合个数等条件进行优化，成功对 NEAT1 成像。

该结构结合生化实验检测Cas13b的稳定性、动力学和功能，为Cas13b靶RNA的识别提供了一个机制模型，并确定了目标特异性和切割特异性的特征。 在这些观察的基础上，研究产生了具有改变的切割偏好的Cas13b变异体。

Using Cas13, we developed a programmable RNA editing system that allows for temporal modulation of genetic variants in transcripts. The system, called RNA Editing for Programmable A to I Replacement (REPAIR), works by fusing the adenosine deaminases acting on RNA (ADAR2) deaminase domain to Cas13b (Fig. 3a). Because the optimal substrate for ...

2023年9月18日 · 目前有3种基于CRISPR-Cas13系统的RNA成像工具：Broad研究所张锋团队研发的dLwaCas13a-msfGFP融合蛋白、用于LncRNA失踪的dPspCas13b-EGFP、进行活体转录本成像的CRISPR Live FISH技术。 3、转录组调控：在dCas13蛋白上融合各种效应蛋白，能够实现各种针对RNA的调控： m6A甲基化调控，如dCas13-METTL3和dCas13-METTL3/METTL14融合蛋白，可实现特定位点的RNA甲基化。 dCas13-ALKBH5融合蛋白，实现特定位点RNA去甲基化。 …

Through a combination of biochemical and genetic experiments, we show that Cas13b processes its own CRISPR array with short and long direct repeats, cleaves target RNA, and exhibits collateral RNase activity.

2024年5月3日 · Smargon, A. A. et al. Cas13b is a type VI-B CRISPR-associated RNA-guided RNase differentially regulated by accessory proteins Csx27 and Csx28. Mol. Cell 65, 618–630 (2017).

2021年8月30日 · CRISPR–Cas13 systems have been developed for precise RNA editing, and can potentially be used therapeutically when temporary changes are desirable or when DNA editing is challenging. We have...

2017年2月16日 · Using a computational sequence database mining approach, we identify two class 2 CRISPR-Cas systems (subtype VI-B) that lack Cas1 and Cas2 and encompass a single large effector protein, Cas13b, along with one of two previously uncharacterized associated proteins, Csx27 and Csx28.

2018年11月13日 · To investigate how Cas13b processes pre-crRNA, recognizes crRNA and settles the spacer nucleotides for target recognition, we solved the crystal structure of Bergeyella zoohelcum Cas13b...