Feb 11, 2025 · We show that collateral activity only occurs with high Rfx Cas13d expression. Using low-copy Rfx Cas13d in transcriptome-scale and combinatorial pooled screens, we achieve high on-target...

Aug 11, 2022 · 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...

Apr 12, 2024 · 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...

Feb 21, 2024 · 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 sequence-specific degradation without ...

Mar 31, 2022 · 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 ...

Apr 19, 2018 · 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.

Learn about new RNA-targeting Cas13d enzymes that average only 2.8 kb in size, are easy to package in low-capacity vectors, have high on-target and low off-target editing rates.

Mar 30, 2022 · The Cas13d assembled with pre-crRNA array and cleaves within the crRNA to form mature crRNA consisting of 30 nucleotides, 5′ direct repeat (DR) region, and 20 nucleotides, 3′ spacer region that is complementary to the target ssRNA.

Dec 12, 2023 · A large-scale screen of CRISPR-Cas13d guide sequences is used to build a deep learning model to predict high-efficiency guides for both CasRx and a new effector, DjCas13d, that enables highly efficient RNA targeting with minimal cellular toxicity and off-target effects.

May 5, 2020 · Cas13d, a subtype of Cas13, has been used for efficient RNA knockdown in mammalian cells. Yet little is known about the design rules of guide RNA (gRNA) for optimal Cas13d activity.