KaryoCreate: A CRISPR-Based Technology to Study Chromosome-Specific Aneuploidy
- Description
This study described karyotype CRISPR engineered aneuploidy technology (KaryoCreate), a system that enables generation of chromosome-specific aneuploidies by co-expression of an sgRNA targeting chromosome-specific CENPA-binding ɑ-satellite repeats together with dCas9 fused to mutant KNL1. They designed unique and highly specific sgRNAs for 19 of the 24 chromosomes. To examine the activity of the predicted sgRNAs, they co-expressed selected sgRNAs with Cas9 and monitored cell proliferation. For the study, they used hTERT TP53−/− human colonic epithelial cells (hCECs) and hTERT TP53 WT retinal pigment epithelial cells (RPEs) expressing p21 (CDKN1A) and RB (RB1) shRNAs. Transduced Cas9-expressing RPEs and hCECs with a lentiviral vector expressing either a centromeric or a negative control sgRNA (sgNC) that does not target the human genome. The dataset includes single-cell RNA sequencing and bulk RNA sequencing data.
Access
- Restrictions
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Free to All
- Instructions
- Single-cell and bulk RNA sequencing data have been deposited at Gene Expression Omnibus (GEO) and all original analysis code has been deposited to GitHub.
- Grant Support
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MRA Young Investigator Award/Tara Miller Melanoma FoundationCancer Grand Challenges/Cancer Research UKC5470/A27144/Mark Foundation for Cancer ResearchGR-2018-12367077/Ricerca FinalizzataFondazione Cariplo/Fondazione CariploAIRC-MFAG 2018 ID.21665/Italian Association for Cancer ResearchAIRC Start-Up 2020 ID.25189/Italian Association for Cancer Research