S-phase Increases CRISPR/Cas9 and ssODN Directed Editing.
2015-06-08T02:45:41Z (GMT) by
<p>HCT116-19 cells were seeded at 2.5x10<sup>6</sup> cells in a 100mm dish and synchronized for 24 hours with 6uM aphidicolin then released for 4 hours. Synchronized and unsynchronized cells were electroporated at a concentration of 5x10<sup>5</sup> cells/100ul with CRISPR/Cas9 and 72NT ssODN under the standard reaction conditions. Following electroporation, cells were seeded in 6-well plates and allowed to recover for 48 hours before flow cytometry analysis was carried out. Correction efficiency (%) was determined by the number of viable eGFP+ cells. Each sample set was performed in duplicate and ± represent calculated standard deviation per sample. The unsynchronized data is the same shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0129308#pone.0129308.g004" target="_blank">Fig 4</a>. Statistical analysis was performed using two-sample unequal variance students T-test distribution to compare the value of correction efficiency between synchronized and un-synchronized cells when treated with CRISPR/Cas9.</p><p>*p<0.05</p><p>S-phase Increases CRISPR/Cas9 and ssODN Directed Editing.</p>