# Delivery Technologies for In Vivo Genome Editing

> **NIH NIH UH3** · BETH ISRAEL DEACONESS MEDICAL CENTER · 2022 · $1,385,636

## Abstract

Project Summary
 New in vivo delivery technologies are urgently needed that enable selective genome editing of somatic
cells without the limitations of existing viral delivery systems or lipid nanoparticles. We propose to develop two
complementary strategies. First, by tethering Cas9 and base editor ribonucleoproteins (RNPs) to homing
moieties, such as antibodies or nucleic acid aptamers, we will develop delivery systems capable of editing a
specific population of target cells. As a second approach, we will engineer viral like particles (VLPs) to facilitate
efficient, tissue and cell specific delivery of genome editing agents. In the process, we will develop delivery
systems that are capable of targeting hematopoietic stem and progenitor cells (HSPCs), among other tissues.
To evaluate the efficiency and cell-type specificity of our proposed delivery methods, we will also generate a
reporter mouse that quantitatively and sensitively reports genome editing from base editors or programmable
nucleases without requiring DNA sequencing. In this proposal, we intend to:
(1) Design targeted ribonucleoprotein conjugates that selectively bind, enter, and edit target cells. Cell
and tissue selective Cas9 and base editor RNP delivery systems will be designed by tethering genome editing
proteins, directly or indirectly, to aptamer and antibody targeting moieties. The kinetics, magnitude, and
specificity of RNP endocytosis, endosomal escape, and nuclear transport will be defined and genome editing
efficiency and targeting specificity determined in vitro and in vivo.
(2) Engineer ribonucleoprotein nanoparticle delivery systems for cell and tissue targeted genome
editing. SV40 capsid proteins will be engineered to form viral like particles (VLPs) that are capable of
packaging ribonucleoproteins, rather than DNA. The stoichiometry of VLP-RNP delivery systems, which affords
optimal cell uptake, endosomal escape, and nuclear transport will be defined. Targeting specificity, as
determined by viral capsid tropism will be defined, and genome editing efficiency analyzed in vitro and in vivo.
(3) Develop a reporter mouse for facile assessment of targeted genome editing efficiency and cell- and
tissue-type specificity. We will optimize a reporter gene to independently detect base editing, end-joining, or
homology-directed repair. The reporter will be integrated into the Rosa26 safe harbor locus in C57BL/6 mouse
embryonic stem cells to generate transgenic mice. Genome editing outcomes will be evaluated by
fluorescence and luminescence measurements and correlated with high throughput DNA sequencing.
(4) Demonstrate safe and effective delivery of genome editing agents in non-human primates. The
delivery of genome editors to HSPCs and other target tissues will be assessed in rhesus macaques. Both
mammalian and non-mammalian systems will be evaluated to optimize large scale production of the genome
editor and related RNP delivery components. Targeting specificity and genome ed...

## Key facts

- **NIH application ID:** 10664097
- **Project number:** 4UH3AI150551-04
- **Recipient organization:** BETH ISRAEL DEACONESS MEDICAL CENTER
- **Principal Investigator:** Elliot Chaikof
- **Activity code:** UH3 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $1,385,636
- **Award type:** 4N
- **Project period:** 2019-08-22 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10664097

## Citation

> US National Institutes of Health, RePORTER application 10664097, Delivery Technologies for In Vivo Genome Editing (4UH3AI150551-04). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10664097. Licensed CC0.

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