Identification of genes involved in cell cycle regulation using arrayed synthetic CRISPR RNA libraries in a multiparameter high-content assay
Poster Apr 18, 2018
Žaklina Strezoska, Matthew R. Perkett, Eldon T. Chou, Elena Maksimova, Emily M. Anderson, Shawn McClelland, Melissa L. Kelley, Annaleen Vermeulen and Anja van Brabant Smith
Here, we used a cell cycle reporter cell line to perform an arrayed, synthetic crRNA:tracrRNA screen targeting 169 genes with four crRNAs per gene with high content analysis (HCA) to identify genes that regulate the cell cycle. Multiple parameters were used to classify cells into different cellular states and phases of the cell cycle: cells with irregularly shaped nuclei, cells in G1 phase, cells either in S or G2 phase, cells in mitosis or with condensed chromatin, and cells with multinuclear DNA component. We used a novel statistical method for hit identification and applied multiple strategies including gene expression analysis, confirmation of genomic insertions and deletions, and validation by orthogonal reagents to identify high confidence target genes with roles in cell cycle regulation. Most hits had multiple positive crRNAs per target gene, enabling identification of target genes with high confidence, demonstrating the power of combining synthetic crRNAs libraries with HCA assays in screening for complex cellular phenotypes in an arrayed format. Given the ease of transfecting RNAs into most cell types, we expect many phenotypic assays to be amenable to arrayed screening with synthetic crRNA libraries.
Psychiatric Risk Gene Cacna1c and Early Life Stress: Potential Gene-Environment interactions?Poster
Early life stress (ELS) is highly associated with development of psychopathology
and mood disorders in adulthood. Genetic studies have identified variation in the gene calcium voltage-gated channel subunit alpha1C (CACNA1C) to increase risk for several psychiatric disorders. This poster assessed the expression of Cacna1c following prepubertal stress.
Treatment Options for Chronic Parvovirus Viremia in Pediatric Heart Transplant Patients in a Tertiary Care CenterPoster
This abstract discusses three cases of pediatric heart transplant patients who suffered from parvovirus (B19) infection. Of these patients, two ( B & C) responded well to standard intravenous Ig therapy. Patient A however, did not respond to standard treatment and was begun on subcutaneous Ig, which effectively diminished his viral load. Thus, subcutaneous Ig infusions might serve as a second line treatment for transplant patients with parvovirus who do not respond well to the standard approach.READ MORE
T-helper cell phenotype expression in cutaneous lesions of angioimmunoblastic T-cell lymphomaPoster
Angioimmunoblastic T-cell lymphoma (AITL) is a common type of peripheral T-cell lymphoma. AITL can be missed until lymphadenopathy develops in patients initially presenting with skin lesions, as skin biopsy may lack conclusive findings. Our case highlights the extranodal presentation of AITL with cutaneous lesions displaying the TFH phenotype.READ MORE