KaryoCreate: A CRISPR-based technology to study chromosome-specific aneuploidy by targeting human centromeres

Area of Science:

• Genetics and Genomics
• Cancer Biology
• Molecular Biology

Background:

• Aneuploidy, characterized by chromosome number alterations, is a common feature in cancer cells.
• Understanding the functional consequences of specific aneuploidies is crucial for cancer research.

Purpose of the Study:

• To introduce KaryoCreate, a novel CRISPR-engineered system for generating chromosome-specific aneuploidies.
• To validate the efficiency and specificity of KaryoCreate in inducing chromosome missegregation.
• To investigate the role of specific aneuploidies in cancer progression, using colon epithelial cells as a model.

Main Methods:

• KaryoCreate utilizes co-expression of chromosome-specific sgRNAs and dCas9 fused to mutant KNL1.
• sgRNAs target CENPA-binding alpha-satellite repeats for chromosome-specific missegregation.
• CRISPR-based system validated in cellular models to induce chromosome gains and losses.

Main Results:

• KaryoCreate successfully generated chromosome-specific aneuploidies with an average efficiency of 8% for gains and 12% for losses.
• The technology was validated across 10 different chromosomes.
• Chromosome 18q loss in colon cells induced resistance to TGF-β, linked to hemizygous gene deletions.

Conclusions:

• KaryoCreate is an innovative technology for creating and studying chromosome missegregation and aneuploidy.
• This system provides a powerful tool for investigating the role of aneuploidy in cancer and other biological contexts.
• Specific aneuploidies, like 18q loss, can drive cancer phenotypes such as drug resistance.