Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain
the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in
Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles
and JavaScript.
Recent advances in genome editing technologies have substantially improved our ability to make precise changes in the genomes of eukaryotic cells. Programmable nucleases, particularly the CRISPR/Cas system, are revolutionizing our ability to interrogate the function of the genome and can potentially be used clinically to correct or introduce genetic mutations to treat diseases that are refractory to traditional therapies. This collection of recent articles from the Nature Research journals provides an overview of current progress in developing targeted genome editing technologies. A selection of protocols for using and adapting these tools in your own lab is also included.
The authors introduce MACHETE (molecular alteration of chromosomes with engineered tandem elements), a clustered regularly interspaced short palindromic repeats directed Cas9-based system for the efficient deletion of megabase-sized genomic regions.
This protocol provides extensive guidelines and detailed steps to generate novel bio-engineered bacterial strains using CRISPR-associated transposase (CAST) systems, with available plasmids and standard molecular biology techniques.
The authors provide an expanded CRISPR–Cas9-assisted recombineering toolkit for rapid and efficient engineering of genetically intractable Pseudomonas aeruginosa isolates.
The authors describe a new base editing system—the transformer base editor—to induce efficient editing with no observable genome-wide or transcriptome-wide off-target mutations, both in mammalian cells and in mice.
This protocol for universal and proficient Pseudomonas recombineering uses phage-encoded homologous recombination–Cas3 systems, including SacB counterselection and Cre site-specific recombinase for two- or three-step seamless genome modification.
The authors provide a protocol for cytosine base editing to introduce precise substitutions into the genome of zebrafish, an important model for genetic studies and in vivo disease modeling.
Detect-seq (dU-detection enabled by C-to-T transition during sequencing) enables off-target evaluation of programmable cytosine base editors. DNA is chemically labeled, and the editing intermediate dU is enriched for sequencing by biotin pull-down.
This CRISPR-Combo system enables efficient multiplexed orthogonal genome editing and transcriptional activation in plants. Here, the use of CRISPR-Combo is demonstrated for speed breeding of transgene-free, genome-edited Arabidopsis and enhancing rice regeneration with more germline mutations.
This protocol for Multiplexed Intermixed CRISPR Droplets uses microfluidics to create droplets containing Cas9, multiplexed single-guide RNAs and corresponding DNA barcodes, allowing large-scale genetic screens to be performed in F0 zebrafish.