Evaluation of Gene-Corrected Hematopoietic Stem Cells for Treating Sickle Cell Disease

PI’s: David Archer

Specific Aim 1: Delivery of fluorescent proteins into single HSCs via microinjection and evaluate their functionality. We will use a novel microfluidics-based approach for single-cell injection to optimize delivery conditions and single cell sorting, increase throughput, and evaluate the viability and differentiability of injected HSCs. The fluorescent protein mCherry will be used as a model protein as well as a marker for injected cells.

Specific Aim 2: Delivery of ZFNs or TALENs into single HSCs from sickle cell mice and evaluate their functionality. We will deliver different amounts of ZFNs or TALENs into HSCs and evaluate the functionality of injected HSCs using a single cell liquid culture/RBC expansion assay. We will also validate the efficacy of transplant assays for single HSCs.


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