TL;DR
This engineering method allows for endogenous expression of the Soc-mCherry recombinant protein within Escherichia coli cells during phage replication, after which the recombinant protein spontaneously assembles onto the capsid of the engineered phage.
A one-step CRISPR-based transformation protocol developed in this study enables fast genome editing workflows with minimal hands-on time and demonstrates the versatility of the modified system for this industrially important genus.
Cas9 ablation enabled restoration of CUL4B expression and function following introduction of a CUL4B expression plasmid, providing proof of concept for a broadly applicable approach to studying protein function through rescue experiments.
An RNA-guided bridge recombinase system is engineered through rational mutagenesis and AI-assisted directed evolution, enabling programmable chromosomal rearrangements in both plant and mammalian cells and achieving up to a 29.8-fold increase in activity.
A detailed protocol for the production of high-titer, third-generation LVs with a dual-expression cassette for Cas13d and a customizable guide RNA is described, which can be used for stable transduction and efficient RNA knockdown in a broad range of mammalian cell types.
Genome editing in avian systems has largely relied on plasmid-based CRISPR systems and antibiotic selection to achieve successful editing. However, plasmid delivery can result in prolonged nuclease expression and potential unintended DNA integration. In addition, selection-based enrichment may mask the intrinsic geno...
Summary Programmable DNA integration using CRISPR-associated transposase elements (CASTs) offer powerful capabilities for genome engineering. The large single effector Cas12k CAST examples evolved from a minimal TnpB nuclease protein. Here, we engineer a de novo RNA-guided transposition systems in bacteria, where the s...
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