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Packaged delivery of CRISPR-Cas9 ribonucleoproteins accelerates genome editing.
Nucleic Acids Res
February 2025
Department of Chemistry, University of California, Berkeley, CA 94720, United States.
Effective genome editing requires a sufficient dose of CRISPR-Cas9 ribonucleoproteins (RNPs) to enter the target cell while minimizing immune responses, off-target editing, and cytotoxicity. Clinical use of Cas9 RNPs currently entails electroporation into cells ex vivo, but no systematic comparison of this method to packaged RNP delivery has been made. Here we compared two delivery strategies, electroporation and enveloped delivery vehicles (EDVs), to investigate the Cas9 dosage requirements for genome editing.
View Article and Find Full Text PDFDissecting tumor cell programs through group biology estimation in clinical single-cell transcriptomics.
Nat Commun
March 2025
Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA.
With the growth of clinical cancer single-cell RNA sequencing studies, robust differential expression methods for case/control analyses (e.g., treatment responders vs.
View Article and Find Full Text PDFNanoscale Biodegradable Printing for Designed Tuneability of Vaccine Delivery Kinetics.
Adv Mater
February 2025
Department of Materials, Department of Bioengineering, Institute of Biomedical Engineering, Imperial College London, South Kensington Campus, London, SW7 2AZ, United Kingdom.
Two-photon polymerization (2PP) 3D printing enables top-down biomaterial synthesis with nanoscale spatial resolution for de novo design of monodisperse injectable drug delivery systems. Spatiotemporal Controlled Release Inks of Biocompatible polyEsters (SCRIBE) is a novel poly(lactic-co-glycolic acid)-triacrylate resin family with sub-micron resolution and tuneable hydrolysis that addresses the limitations of current 2PP resins. SCRIBE enables the direct printing of hollow microparticles with tuneable chemistry and complex geometries inaccessible to molding techniques, which are used to engineer controlled protein release in vitro and in vivo.
View Article and Find Full Text PDFCRISPRi-ART enables functional genomics of diverse bacteriophages using RNA-binding dCas13d.
Nat Microbiol
March 2025
Innovative Genomics Institute, University of California, Berkeley, CA, USA.
Bacteriophages constitute one of the largest reservoirs of genes of unknown function in the biosphere. Even in well-characterized phages, the functions of most genes remain unknown. Experimental approaches to study phage gene fitness and function at genome scale are lacking, partly because phages subvert many modern functional genomics tools.
View Article and Find Full Text PDFLatent activity in TnpB revealed by mutational scanning.
bioRxiv
February 2025
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, 94720, USA.
TnpB is an evolutionarily diverse family of RNA-guided endonucleases associated with prokaryotic transposons. Due to their small size and putative evolutionary relationship to Cas12s, TnpB holds significant potential for genome editing and mechanistic exploration. However, most TnpBs lack robust gene-editing activity, and unbiased profiling of mutational effects on editing activity has not been experimentally explored.
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