Fig. 2
From: Pooled clone collections by multiplexed CRISPR-Cas12a-assisted gene tagging in yeast
CRISPR-Cas12a (Cpf1)-assisted tag library engineering (CASTLING) in a nutshell. a For each target locus, a DNA oligonucleotide with site-specific homology arms (HAs) and a CRISPR spacer encoding a target-specific CRISPR RNAs (crRNA) is designed and synthesized as part of an oligonucleotide array. The resulting oligonucleotide pool is recombineered with a custom-tailored feature cassette into a pool of self-integrating cassettes (SICs). This results in a clone collection (library) that can be subjected to phenotypic screening and genotyping, for example, using Anchor-Seq12. b The three-step recombineering procedure for SIC pool generation; details are given in the main text and Methods
