| Description |
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CRISPR/Cas9
ribonucleoprotein (RNP) complexes offer a transient way of precise genome editing via
homology-directed repair (HDR) with minimal potential off-target effect. However,
intracellular delivery of RNP and donor templates remains challenging. This thesis
reports a delivery vehicle based on cationic lipidoid nanoparticles (LNP) that can
simultaneously deliver Cas9 RNP and a repair ... read moretemplate into a fluorescence reporter cell
line and subsequently induce CRISPR-mediated HDR. More than 100 kinds of LNPs were
formulated and screened for their efficacy on delivery of Cas9 RNP and ssODN. Positive
results were observed via fluorescence microscope and flow cytometry. These LNPs were
further optimized by varying formulation and cargo/LNP ratio. 8 % - 16 % HDR efficiency
was reached under different delivery conditions. MTT assay was performed and showed our
LNPs had low toxicity. These result shows that lipidoid nanoparticles loaded with CRISPR
components can generate HDR editing efficiently and have tremendous potential for
treating genetic diseases.
Thesis (M.S.)--Tufts
University, 2019.
Submitted to the Dept. of Chemical
and Biological Engineering.
Advisor: Qiaobing
Xu.
Committee: Emmanuel Tzanakakis, and Xiaocheng
Jiang.
Keyword:
Bioengineering.read less
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