AI Networking Cookbook: Practical recipes for AI-assisted network automation and development
TL;DR
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.
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.
Kilobase-scale nickase-targeting (KNIT) editing enables single-nick-based DNA insertion across genomic loci and cell types, supporting insertion of DNA fragments exceeding 10 kb, with high efficiency and minimal unwanted off-target effects.
MUTATOR is reported, a MUlTiplexAble and self-iTerative ORthogonal base-editing platform that enables N-to-N diversification in Escherichia coli and is established as a broadly applicable platform for genome-wide diversification, functional dissection, and rapid engineering of industrial microbial chassis.
A practical protocol for using either PE or iPE to introduce point mutation(s) or small-to-medium sized insertions and deletions in cell culture, and how to assess editing efficiency via flow cytometry and next-generation sequencing is provided.
This review compares Cas9-mediated homology-directed repair (HDR) with generations of cytosine base editors (CBE1–CBE3), adenine base editors (ABE1-ABE7), and prime editors (PE1–PE3b), focusing on their mechanistic distinctions, efficiencies, delivery challenges, and therapeutic applications.
Base editors hold great promise in endogenous mutagenesis for genetic screening. However, the development of base editors that induce saturated multi-base conversions with diverse mutation spectrum is challenging. Here, we develop triple base editors (smACGs) that simultaneously mutagenize adenine, cytosine, and guanin...
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