In vitro gene editing of CD34 + hematopoietic stem cells and progenitor cells (HSPCs) provides a great opportunity to develop new therapies for a wide range of malignant and non-malignant diseases. Currently, delivering CRISPR using electroporation and/or viral transduction has enabled effective gene editing in HSPCs, but cytotoxicity is a drawback of current methods of use. Nanoparticle (NP) based gene editing strategies can further enhance the gene editing potential of HSPCs and provide a delivery system for in vivo applications. Here, Christina Eich et al. from Leiden University Medical Center in the Netherlands developed CRISPR/ Cas9-plga-NP which can effectively encapsulate Cas9 protein, single gRNA and fluorescent probes.












Main points of this article:



1) The initial "burst" of Cas9 and gRNA release is followed by a sustained release pattern.



2) CRISPR/ Cas9-plga-NP was absorbed and processed by human HSPCs without causing cytotoxicity.














3) After escape from lysosomal compartment, CRISPR/ Cas9-plga-NPS-mediated gene editing of gamma-globin loci leads to increased expression of fetal hemoglobin (HbF) in primary erythroid cells.







Taken together, the development of CRISPR/ Cas9-plga-NP provides an attractive tool for delivering THE CRISPR component to targeted HSPCs and could provide the basis for in vivo treatment of hemoglobin and other genetic diseases.














reference



Luis J. Cruz, et al. PLGA-Nanoparticles for Intracellular Delivery of the CRISPR-Complex to Elevate Fetal Globin Expression in Erythroid Cells. Biomaterials, 2020.DOI: https://doi.org/10.1016/j.biomaterials.2020.120580https://doi.org/10.1016/j.biomaterials.2020.120580





Source: Theory of Oddities



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