OCT 03, 2018 6:00 AM PDT

Identification of Pre-existing Adaptive Immune Responses to Cas9 in Humans

Presented at: CRISPR 2018
C.E. Credits: P.A.C.E. CE Florida CE
Speaker
  • PhD Student in Stem Cell Biology and Regenerative Medicine, Stanford University
    Biography
      Dr. Charlesworth grew up in Australia and moved to the US in 2008. He went to Humboldt State for undergrad where he obtained his bachelors in molecular biology & zoology in 2015. After completing his bachelors he started a CIRM Bridges stem cell internship at Stanford in 2015 in the lab of Dr. Matthew Porteus where he worked on the development of genome editing in Hematopoietic Stem Cells and identified the presence of pre-existing adaptive immune responses to Cas9 in humans. In September of 2017 he started a PhD in Stem Cell and Regenerative Medicine at Stanford and is now a graduate student in the Nakauchi lab.

    Abstract

    The CRISPR-Cas9 system has proven to be a powerful tool for genome editing allowing for the precise modification of specific DNA sequences within a cell. Many efforts are currently underway to use the CRISPR-Cas9 system for the therapeutic correction of human genetic diseases. The most widely used homologs of the Cas9 protein are derived from the bacteria Staphylococcus aureus (S. aureus) and Streptococcus pyogenes (S. pyogenes). Based on the fact that these two bacterial species cause infections in the human population at high frequencies, we looked for the presence of pre-existing adaptive immune responses to their respective Cas9 homologs, SaCas9 (S. aureus homolog of Cas9) and SpCas9 (S. pyogenes homolog of Cas9). We have identified the presence of pre-existing humoral immune responses to SaCas9 and SpCas9 by both Western Blot and ELISA. In addition we have also identified pre-existing cellular responses to Cas9 by intracellular cytokine staining and ELISPOT. 

    Learning Objectives: 

    1. Pre-existing adaptive immunity to the most commonly used homologous of Cas9 is prevalent
    2. Potential immune responses to any new therapeutic should be considered when considering any kind of in-vivo therapy


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