Resources

CRISPR/Cas9 technology was used on cells to study JC polyomavirus that causes progressive multifocal leukoencephalopathy. CRISPR/Cas9 construct was delivered to the cells to randomly introduce stop codons in the gene for each entry receptor for the pathogen. Single cell isolation and expansion was performed to obtain a clonal population using CellRaft AIR System. Deep sequencing of the CRISPR editing was performed at the Center for Computational and Integrative Biology (CCIB) DNA core at the Massachusetts General Hospital (MGH). Clones displaying mutations of interest were selected for further investigation and discovered that the second intracellular loop of each receptor was necessary for infection. Using the CellRaft AIR System to identify and expand clones, a better understanding of viral infection was identified in addition to understanding normal function of these receptors for serotonin.