PREditOR: a synthetic biology approach to removing heterochromatin from cells

• Published in: Chromosome research Vol. 24; no. 4; pp. 495 - 509
• Main Authors: Molina, Oscar, Carmena, Mar, Maudlin, Isabella E., Earnshaw, William C.
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.12.2016; Springer Nature B.V
• Subjects: Animal Genetics and Genomics; Biomedical and Life Sciences; Cell Biology; Cell division; Cell Line; Centromere - metabolism; Chromatin; Chromosome Segregation; Chromosomes; drugs; enzyme inhibitors; enzymes; gene targeting; genes; Genomic Instability; heterochromatin; Heterochromatin - physiology; Human Genetics; Humans; Life Sciences; Mitosis; Original; Original Article; Plant Genetics and Genomics; Synthetic biology; Synthetic Biology - methods; Heterochromatin; Mitosis; Chromosome segregation; Centromeres
• ISSN: 0967-3849; 1573-6849; 1573-6849
• DOI: 10.1007/s10577-016-9539-3

It is widely accepted that heterochromatin is necessary to maintain genomic stability. However, direct experimental evidence supporting this is slim. Previous studies using either enzyme inhibitors, gene knockout or knockdown studies all are subject to the caveat that drugs may have off-target effects and enzymes that modify chromatin proteins to support heterochromatin formation may also have numerous other cellular targets as well. Here, we describe PREditOR (protein reading and editing of residues), a synthetic biology approach that allows us to directly remove heterochromatin from cells without either drugs or global interference with gene function. We find that removal of heterochromatin perturbs mitotic progression and causes a dramatic increase in chromosome segregation defects, possibly as a result of interfering with the normal centromeric localization of the chromosomal passenger complex.