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A phage-encoded small RNA that mimics chimeric guide to inhibit CRISPR-Cas9

CRISPR Cas9 relies on a dual crRNA-tracrRNA guide, yet whether RNA based anti CRISPRs exist for this system has remained unknown. Here we identify a phage encoded small non coding RNA, rAcrIIA1, that adopts an unexpected chimeric crRNA-tracrRNA architecture, faithfully recapitulating the entire single guide RNA (sgRNA) of Cas9. Cryo EM structures of the Cas9-rAcrIIA1 binary complex and the Cas9-rAcrIIA1-DNA ternary complex reveal that rAcrIIA1 engages Cas9 nearly identically to the native...

bioRxiv 7d ago

dCas allele sequestration (das-CRISPR): A Versatile New Method to Achieve Monoallelic Gene Editing in Mouse Embryos and in cell culture.

CRISPR-Cas9 technology is a powerful tool extensively used for genome editing in mouse and many other species. Streptococcus pyogenes Cas9 efficiently cuts both alleles in mouse zygotes leaving many edited embryos without a functional protein that might be needed to sustain development, to survive postnatally or to reproduce, thus complicating its overwhelmingly advantageous use in making gene modifications. About 25% of mouse genes are essential for embryonic development and another 7% are...

bioRxiv 6d ago

An improved CRISPR-base editor tool to target virulence factors in the ruminant pathogen Mycoplasma bovis

Mycoplasma bovis is a minimal bacterium infecting cattle, which causes a wide variety of symptoms and is impacting dairy and beef producers worldwide. Part of the difficulty in research surrounding M. bovis, and other mycoplasmas, is the lack of efficient genome editing tools. We previously presented a proof of concept of a transposon-based CRISPR-Base Editor system to introduce targeted mutations in M. bovis.

bioRxiv 11d ago

CRISPR-mediated Stxbp1 gene activation ameliorates epileptic and aggressive phenotypes in Stxbp1-haploinsufficient mice

Mutations in the syntaxin-binding protein 1 (STXBP1) gene, which encodes the presynaptic protein Munc18-1, cause a spectrum of severe epileptic encephalopathies and neurodevelopmental disorders, including Ohtahara syndrome, for which no curative treatment is currently available. Because the disease pathomechanism is thought to be driven by haploinsufficiency, restoring expression of the wild-type allele to physiological levels could provide therapeutic benefit. Here, we evaluated...

bioRxiv 1d ago

S-8 - CRISPR Therapeutics AG (0001674416) (Filer)

Filed: 2026-06-05 AccNo: 0001193125-26-258473 Size: 387 KB

SEC EDGAR Filings 5d ago

Multiple approaches for CRISPR-based targeting of DNA methylation to promoters of bacterial and viral susceptibility genes in cassava

Targeted epigenetic modifications of specific gene regulatory regions have the potential to confer beneficial traits for crop improvement. Two recently developed CRISPR/Cas9-based epigenome editing tools were tested in transgenic cassava to target cytosine methylation to the promoter region of MeSWEET10a, a necessary gene for infection by the Cassava bacterial blight pathogen, Xanthomonas phaseoli pv. manihotis. The two systems leverage unique methyltransferases, and each induced distinct...

bioRxiv 4d ago

Are we getting to the point where it's safe to gene-edit babies?

When a rogue researcher in China revealed in 2018 that he had used CRISPR to create three gene-edited children, his actions were almost universally condemned by biologists around the world. The main objection was not that gene-editing babies is wrong in itself, but that the CRISPR technique used was not safe and had a very high risk of causing harmful mutations. Now, a team in the US has used an improved form of CRISPR, known as base editing, to edit healthy embryos and shown that it can be...

New Scientist 4d ago

DNA remodeling couples target recognition to directional transposition in a Tn7-like CAST

CRISPR-associated transposons (CASTs) couple target recognition to the insertion of large DNA cargoes, but how distinct targeting pathways are converted into productive and directional integration remains poorly understood1. Here we define the assembly pathway of a type I-B1 CAST from Anabaena variabilis, a system closely related to prototypical Tn7 that retains TnsD-mediated glmS recognition while incorporating CRISPR-based RNA-guided targeting2. Cryo-electron microscopy structures across...

bioRxiv 7d ago

New 'SMArT' platform makes gene editing in hematopoietic stem cells more efficient and safer

New 'SMArT' platform makes gene editing in hematopoietic stem cells more efficient and safer Sadie Harley Scientific Editor Robert Egan Associate Editor A team of researchers led by Luigi Naldini at the San Raffaele Telethon Institute for Gene Therapy (SR-Tiget) has developed a new strategy to significantly improve the precision and safety of CRISPR-Cas9 gene editing in human blood stem cells, potentially overcoming one of the major barriers limiting broader clinical application of genome...

Phys.org 9d ago

Fusion of a non-specific DNA-binding domain enhances Cas12a trans-cleavage for robust nucleic-acid diagnostics

CRISPR-Cas12a underlies powerful genome-editing and nucleic-acid detection technologies, yet its performance is limited by inefficient target engagement and low catalytic turnover, particularly at low target abundance and elevated temperatures. Here, we report a modular protein-engineering strategy to enhance Cas12a trans-activity by fusing the hyperthermophilic DNA-binding protein Sso7d to the N terminus of Lachnospiraceae bacterium ND2006 Cas12a (LbaCas12a). The resulting fusion enzyme...

bioRxiv 9d ago