DNA double-strand breaks (DSBs) are highly cytotoxic lesions and pose a major threat to genome stability if not properly repaired. We and others have previously shown that a class of DSB-induced small RNAs (diRNAs) is produced from sequences around DSB sites. DiRNAs are associated with Argonaute (Ago) proteins and play an important role in DSB repair, though the mechanism through which they act remains unclear. Here, we report that the role of diRNAs in DSB repair is restricted to repair by homologous recombination (HR) and that it specifically relies on the effector protein Ago2 in mammalian cells. Interestingly, we show that Ago2 forms a complex with Rad51 and that the interaction is enhanced in cells treated with ionizing radiation. We demonstrate that Rad51 accumulation at DSB sites and HR repair depend on catalytic activity and small RNA-binding capability of Ago2. In contrast, DSB resection as well as RPA and Mre11 loading is unaffected by Ago2 or Dicer depletion, suggesting that Ago2 very likely functions directly in mediating Rad51 accumulation at DSBs. Taken together, our findings suggest that guided by diRNAs, Ago2 can promote Rad51 recruitment and/or retention at DSBs to facilitate repair by HR.

Objective To evaluate the effects of cryopreservation in post‐thaw umbilical cord blood units for the survivability of Gram‐positive bacteria strains. Background Microbial screening is required for all cord blood units (CBUs). Four gram‐positive contaminants were documented to survive cryopreservation poorly and isolation of other contaminants were reported. Methods Forty‐eight contaminated CBUs detected with either Staphylococcus epidermidis, Corynebacterium species, Peptostreptococcus or Streptococcus species before cryopreservation were used in this study. CBUs were processed, DMSO‐infused and microbial screened before cryopreservation. Post‐thaw microbial screening was achieved using 1 and 10 ml inoculants in BACTEC culture bottles. Positive bottles were subjected for microbial identification and results were compared with those from pre‐freeze. Results A higher rate of microbial contamination was found using the 10 ml inoculant. Screening of 11 CBUs did not detect any contaminants while 30 CBUs screened detected more than one unknown contaminants and majority of contaminants were identified to be gram‐negative species. Conclusion A higher inoculation volume used at post‐thaw for microbial screening improves contamination detection but leads to the loss of precious cord blood. Some contaminants did not survive cryopreservation or were not identified due to their low microbial levels. Contrasting contaminants found at post‐thaw suggest the improvements made in detection and identification of contaminants over the years.