HiHo-AID2: boosting homozygous knock-in efficiency enables robust generation of human auxin-inducible degron cells.

"rnaseq data was deposited to the ncbi gene entology omnibus database under the accession number: gse243637 ( https://www ncbi nlm nih gov/geo/query/acc cgi?; whole genome sequencing data was deposited to the ncbi sequence readarchive (sra) data repository under the accession number: prjna1050771 ( https://www ncbi nlm nih gov/sra/? clones generated with hdr enhancers (either 1 um m3814 or i53 plus0 25 um m3814) for the 11 targets showed significantly higher homozygous degron-tagging efficiencies (average of 81% varying from 62 to 100%) compared to the 8 targets without hdr enhancers (average of 32% varying from 0 to 83%) (fig 2 h i and additional file 1 : data s1).; the cell clones isolated largely represent bona fide single clones as gpcr of homozygous clones showed no extra band at the wild-type position (additional file 1 : data s1) and functional characterization of several clones at single-cell level provided no indication of contamination from wild-type or heterozygously tagged cells (additional file 1 : fig s5 s7).; additional file 1: note s1-3 fig s1-9 and data s1.; data s1. availability of data and materials vectors enabling hiho-aid2 will be available via addgene (plasmid #216243"

"Declarations Ethics approval and consent to participateNot applicable. Consent for publicationNot applicable. Competing interestsThe authors declare that they have no competing interests. Competing interests The authors declare that they have no competing interests. "

"Funding Open Access funding provided by University of Helsinki (including Helsinki University Central Hospital). This study was supported by Academy of Finland (grants 324929 to EI, 322647 to VMO, 332096 to XZ), Jane and Aatos Erkko Foundation (EI), Foundation Leducq (grant 19CVD04 to EI), Sigrid Juselius Foundation (EI, VMO), EU MSCA-PF (grant 101059424 to MvdS), The Hospital District of Helsinki and Uusimaa/Children and Adolescents (TR), The Paulo Foundation (N.E.), The Maud Kuistila Memorial Foundation (N.E.), and H2020-MSCA-IF (grants 894596 to YW, 813707 to SM)."