🧬 Compact CRISPR nucleases are small — but often slow to activate. A new strategy just fixed that. New preprint from Liu Junjie lab (Tsinghua): fusing ATPase DnaA to PlmCas12e (CasX) via a rigid N-terminal linker overcomes the conformational activation barrier. ⚡ 1.34× average editing boost across 7 endogenous loci in HEK293T 📈 More effective genome targets detected by GenomePAM/GUIDE-seq 🔬 MD simulation: single free-energy basin vs. rugged landscape in WT ❌ ATP hydrolysis mutant: activity drops below wild-type — mechanism confirmed Liu lab, Tsinghua | Preprint doi.org/10.65215/LTSpreprint… #CRISPR #Cas12e #CasX #ProteinEngineering #GeneEditing

🧬 CRISPR evolution — directly inside human cells — for the first time. PNAS: CRISPR-MACE uses adenovirus replication to continuously mutate, select, and amplify Cas9 variants in human cells, bypassing bacterial evolution limitations. Key results: 🔑 "Gatekeeper" G12D mutation: 3× higher DNA affinity 💪 DVKIH 5-mutant variant: ~1000× anti-CRISPR resistance ✅ Validated in CASFISH imaging — brighter, stronger signal Shoulders & Choudhary labs, Broad Institute | PNAS doi.org/10.1073/pnas.2536003… #CRISPR #DirectedEvolution #Cas9 #GeneTherapy #Broadinstitute