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Structure-optimized sgRNA selection with PlatinumCRISPr for efficient Cas9 generation of knockouts.
Genome Res
December 2024
School of Biosciences, College of Life and Environmental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom;
A single guide RNA (sgRNA) directs Cas9 nuclease for gene-specific scission of double-stranded DNA. High Cas9 activity is essential for efficient gene editing to generate gene deletions and gene replacements by homologous recombination. However, cleavage efficiency is below 50% for more than half of randomly selected sgRNA sequences in human cell culture screens or model organisms.
View Article and Find Full Text PDFThe interplay of DNA repair context with target sequence predictably biases Cas9-generated mutations.
Nat Commun
November 2024
Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, UK.
Article Synopsis
- The study investigates how the surrounding DNA sequence affects the repair of double-stranded breaks caused by CRISPR/Cas9, using various genetically modified mouse embryonic stem cell lines.
- Researchers analyzed over 236,000 mutation outcomes from 2800 synthetic DNA sequences, discovering specific roles of DNA repair proteins like Prkdc and Polm in generating small insertions and deletions.
- They developed predictive models for these mutational outcomes based on their findings, enhancing the understanding of DNA repair mechanisms and enabling more accurate control of CRISPR-induced mutations.
How do stretch rate, temperature, and solvent exchange affect elastic network rupture?
Soft Matter
October 2024
School of Polymer Science and Polymer Engineering, University of Akron, Akron, Ohio 44325, USA.
In this study, we investigate three different polymeric networks in terms of their tensile strength as a function of stretching rate, or temperature, or medium viscosity. Both an acrylate-based elastomer and a crosslinked poly(methyl acrylate) are stronger, more stretchable, and tougher at high rates. They are also much stronger at lower temperatures.
View Article and Find Full Text PDFCRISPR-directed gene-editing induces genetic rearrangement within the human globin gene locus.
Gene
December 2024
Gene Editing Institute, ChristianaCare, Delaware 550 South College Avenue, Suite 208, Newark, Delaware 19713, United States. Electronic address:
CRISPR-Cas is a revolutionary technology but has already demonstrated significant feasibility for clinical and non-clinical applications. While the efficiency and precision of this remarkable genetic tool is unprecedented, unfortunately, a series of collateral genetic rearrangement have been reported in response to double-stranded DNA breakage. Once these molecular scissions occur, the cascade of DNA repair reactions can lead to genomic rearrangements especially if breakage takes place within a family of sequence related genes.
View Article and Find Full Text PDFGenome mining for new enediyne antibiotics.
Curr Opin Chem Biol
August 2024
Department of Chemistry, Princeton University, Princeton, NJ 08544, USA; Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA. Electronic address:
Enediyne antibiotics epitomize nature's chemical creativity. They contain intricate molecular architectures that are coupled with potent biological activities involving double-stranded DNA scission. The recent explosion in microbial genome sequences has revealed a large reservoir of novel enediynes.
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