Cas13d：转录组工程中靶向RNA的CRISPR酶 - 知乎 - 知乎专栏
Konermann和Yan等人的新研究描述了新的 Cas13d 酶，其平均大小只有2.8 kb，并且易于包装在低容量载体中!这些小但强大的VI - D型酶是转录组工程中的最新工具。 Cas13d酶是如何被发现的? 微生物CRISPR多样性令人印象深刻，研究者们才刚刚开始挖掘CRISPR的可能性所蕴藏的 ...
Precise RNA targeting with CRISPR–Cas13d - Nature
2025年2月11日 · Using low-copy Rfx Cas13d in transcriptome-scale and combinatorial pooled screens, we achieve high on-target knockdown without extensive collateral activity. Furthermore, analysis of a...
专家点评丨首次证实Cas13d系统在成体动物体内具有靶向沉默RNA …
Cas13蛋白最直接的应用是靶向沉默RNA。目前Cas13a, Cas13b和Cas13d均被证实可以在哺乳动物细胞系中干扰RNA，从而实现基因沉默 【2,3】。 其中，2018年由加州大学伯克利分校Patrick Hsu实验室发现的Cas13d家族蛋白 CasRx 由于体积小，效率高，被认为是在未来应用中最具有优势的Cas13蛋白 【2】。 他们发现与 RNA干扰技术 相比，Cas13d介导的基因沉默具有更高的特异性（与数百个shRNA脱靶相比，Cas13d没有脱靶）和敲除效率（Cas13d达到96％，shRNA …
CRISPR-Cas13系统的分类、功能及应用 - 知乎 - 知乎专栏
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。
A versatile CRISPR-Cas13d platform for multiplexed transcriptomic ...
2024年2月21日 · Cas13d is a compact CRISPR effector with RNA-guided RNA endonuclease activity. 37,38 Unlike Cas9, Cas13d does not bind or cut DNA. 37 Rather, it complexes with a CRISPR-associated RNA (crRNA) containing a programmable ∼23-nucleotide spacer that guides the ribonucleoprotein (RNP) to RNA transcripts for …
High-fidelity Cas13 variants for targeted RNA degradation with
2022年8月11日 · Here, we design a dual-fluorescence reporter system for detecting collateral effects and screening Cas13 variants in mammalian cells. Among over 200 engineered variants, several Cas13 variants...
Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector …
2018年4月19日 · We demonstrated robust target cleavage and collateral RNase activities of the subtype VI-D effectors (Cas13d) along with their ability to process pre-crRNA. Additionally, Cas13d effectors have a strong co-occurrence with polyphyletic WYL-domain-containing accessory proteins.
Cas13d: A New Molecular Scissor for Transcriptome Engineering
2022年3月30日 · To summarize, Cas13d is a “new molecular scissor” that demands more attention to fully unfold its potential and needs an ingenious solution to overcome the associated challenges to become a gold standard in transcriptome editing and application beyond RNA editing with salutary results in the coming future. Author Contributions
Engineered, nucleocytoplasmic shuttling Cas13d enables highly …
2024年4月12日 · Cas13d-NCS transfers nuclear crRNAs to the cytosol, where the protein/crRNA complex binds and degrades complementary target RNAs. We screened various designs of shuttling proteins and...
Cas13d: A New Molecular Scissor for Transcriptome Engineering
2022年3月31日 · Therefore considering the advantages of Cas13d over previously characterized Cas13 subtypes, in this review, we encompass the structural and mechanistic properties of type VI CRISPR-Cas13d systems, an overview of the current reported various applications of Cas13d, and the prospects to improve Cas13d based tools for diagnostic and therapeutic ...