AI Networking Cookbook: Practical recipes for AI-assisted network automation and development
TL;DR
A restriction enzyme-free and ligation-free PCR-based protocol widely applicable to any circular vector that enables targeted mutagenesis of the chikungunya virus (CHIKV) genome, offering a fast, efficient, and reliable strategy for generating mutant constructs suitable for virological and molecular studies.
A programmable gene replacement tool, named prime assembly (PA), which adapts prime editors to produce one or two pairs of 3'-flaps on both the genome and donor DNA, allowing megabase-scale genomic excision and/or kilobase-scale donor insertion at the gene of interest.
A streamlined cloning platform for expressing a gene of interest either alone or fused to a monomeric fluorescent protein under the control of two promoters of choice is described, providing a fast and flexible cloning strategy for protein expression and subcellular localization analyses in mycobacteria.
These findings identify cloning-site proximity to an intact AAV ITR as a major determinant of plasmid stability during bacterial propagation and demonstrate that substantial loss of correctly assembled constructs can occur before AAV production.
This study demonstrates donor-complementary prime editing (DoPE) as a one-step, DSB-free and library-compatible method for precise insertion of large DNA fragments without requiring recombinases or transposases.
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