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© 2021 Noah C. Cheng. This work is freely available courtesy of the author. It may be used under the terms of the Creative Commons Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0) license. For all other uses, please contact the copyright holder.

Creative Commons License

Creative Commons Attribution-No Derivative Works 4.0 International License
This work is licensed under a Creative Commons Attribution-No Derivative Works 4.0 International License.

Degree Name

Bachelor of Arts



First Advisor

Jennifer Hamilton

Second Advisor

Amy Cheng Vollmer


The development of efficient and safe delivery platforms for CRISPR-Cas9 genome editing tools is one of the largest challenges for in vivo therapeutic genome editing. Virus-like particles (VLPs), which harness viral packaging and delivery mechanisms, have been shown to efficiently deliver CRISPR-Cas9 ribonucleoprotein complexes (RNPs) to cells. Here, we sought to elucidate whether VLPs can deliver the necessary components for CRISPR-Cas9-mediated homology-directed repair (HDR). We tested four different methods to induce HDR in a reporter cell line using VLPs: [1] nucleofection of HDR repair templates into cells and subsequent treatment with VLPs carrying Cas9 RNPs (Cas9 VLPs), [2] treatment with Cas9 VLPs loaded with repair templates through electroporation, [3] treatment with VLPs packaging both Cas9 RNPs and a lentiviral genome that encoded the repair template, and [4] treatment with Cas9 VLPs and VLPs carrying just the HDR template encoded on the lentiviral genome (dual-VLP system). We found that VLPs enabled CRISPR-Cas9-mediated HDR, albeit at low frequencies. HDR efficiency was highest when the HDR template was nucleofected into the cells, but appreciable levels of HDR was also observed using the dual-VLP system. This work suggests that VLPs are a viable delivery platform for the necessary components for CRISPR-Cas9- mediated HDR, but considerable improvements in the platform must be made before it is used in therapeutic settings.

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