Researchers at Columbia University used the CellRaft AIR System to support the development of precisely engineered human embryonic stem cell (hESC) lines for a study investigating genetic variants associated with exceptional longevity.
The researchers used CRISPR/Cas9 to introduce two naturally occurring SIRT6 variants enriched in centenarians into the endogenous SIRT6 locus of hESCs. Following gene editing and cell expansion, 10,000 cells were seeded onto a 10,000-microwell CellRaft Array. The CellRaft AIR System was then used to automatically identify and isolate individual colonies and transfer them into 96-well plates for expansion and downstream genotyping.
Expanded clones were screened to identify cells carrying the desired SIRT6 variants and further confirmed by Sanger sequencing. These engineered clonal cell lines were subsequently differentiated into multiple somatic cell types, enabling the researchers to investigate how the centenarian-associated variants affect SIRT6 biology and cellular aging.
The study found that the SIRT6 variants increased endogenous SIRT6 protein levels and helped cells resist replicative and progerin-induced senescence, providing new insight into potential mechanisms underlying healthy aging.
