2023年4月11日 · Through protein and sgRNA engineering, we generate enhanced OsCas12f1 (enOsCas12f1) and enRhCas12f1 variants, with 5’-TTN and 5’-CCD (D = not C) PAMs respectively, exhibiting much higher editing...

2023年7月3日 · Here we present enAsCas12f, an engineered RNA-guided DNA endonuclease up to 11.3-fold more potent than its parent protein, AsCas12f, and one-third of the size of SpCas9. enAsCas12f shows higher DNA...

2021年9月2日 · Cas12f is particularly efficient in the deletion of a pathogenic exon or intron using paired gRNAs with expression individually driven by U6 or H1 promoters.

2023年10月26日 · The type V-F Cas12f from Acidibacillus sulfuroxidans is exceptionally compact (422 amino acids) and has been harnessed as a compact genome-editing tool. Here, we developed an approach, combining deep mutational scanning and structure-informed design, to successfully generate two AsCas12f activity-enhanced (enAsCas12f) variants.

2021年4月19日 · Cas12f, also known as Cas14, is an exceptionally small type V-F CRISPR-Cas nuclease that is roughly half the size of comparable nucleases of this type. To reveal the mechanisms underlying substrate recognition and cleavage, we determined the cryo-EM structures of the Cas12f-sgRNA-target DNA and Cas1 …

2022年9月27日 · Cas12f nucleases are currently one of the most compact CRISPR genome editors. However, the available toolkit of efficient Cas12f editors is limited. Here, we report the characterization and engineering of a miniature CRISPR-Cas12f system from Syntrophomonas palmitatica (SpaCas12f1, 497 amino acids).

2022年9月24日 · In this study, we apply a high-throughput sequencing method to comprehensively assess the editing efficiency, specificity, and safety of four Cas12f proteins in parallel with that of Cas9 and two...

Here, we report a miniature class 2 type V-F CRISPR-Cas genome-editing system from Acidibacillus sulfuroxidans (AsCas12f1, 422 amino acids). AsCas12f1 is an RNA-guided endonuclease that recognizes 5' T-rich protospacer adjacent motifs and creates staggered double-stranded breaks to target DNA.

2021年10月21日 · Our engineered CasMINI variants showed significantly improved gene activation or editing activity over the wild-type Cas12f, suggesting that enhancing interactions in the Cas12f:sgRNA:DNA complex is important for both applications.

Compact Cas proteins, such as Cas12f, should enable the creation of precise genome-editing tools that are easier to deliver with AAVs. Cas12f is particularly efficient in the deletion of a pathogenic exon or intron using paired gRNAs with expression individually driven by U6 or …