# Non-Activated Natural Killer Cell Engineering For Enhanced Cytotoxic Potential

> **NIH NIH R44** · KYTOPEN CORP · 2021 · $300,000

## Abstract

ABSTRACT
Natural killer (NK) cells are a subset of innate immune cells that are able to respond to threat without antibody
priming. This quick response to stimuli makes them an ideal immunotherapy candidate. Yet, genetic modification
in NK cells has proven to be difficult using conventional viral and non-viral transfection methodologies. Alternative
delivery methods are necessary in order to make genetic modifications at reproducible and efficient rates, while
maintaining high cell viability. The proposed study uses continuous flow electric field-assisted transfection via a
proprietary Flowfect™ platform. This platform represents a novel approach to non-viral delivery in historically
“hard-to-transfect” human cells. The current research proposes to transfect non-activated NK cells with Cas9
ribonucleoproteins (RNPs) for genetic modification using the Flowfect™ platform. To achieve this goal, we have
outlined a two-phase research strategy which focuses on stability and functionality of edited NK cells both in vitro
and in vivo. Phase I will focus on optimizing transfection efficiency and maintaining cell functionality while
Phase II will focus on understanding how the Flowfect™ platform would be deployed for pre-clinical Research
and Development purposes. Briefly, our Phase I goal is to 1) determine optimal Flowfect™ conditions in non-
activated NK cells and 2) induce stable knockout of a clinically relevant NK cell target. Meeting our Phase I
milestones will help us build towards our Phase II goals of 1) successfully screening guides which result in
enhanced cytotoxicity and 2) translating these findings to high efficacy within an in vivo mouse model of Acute
Myeloid Leukemia (AML).

## Key facts

- **NIH application ID:** 10139755
- **Project number:** 1R44AI157181-01
- **Recipient organization:** KYTOPEN CORP
- **Principal Investigator:** Bethany Foch Grant
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $300,000
- **Award type:** 1
- **Project period:** 2021-04-20 → 2023-09-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10139755

## Citation

> US National Institutes of Health, RePORTER application 10139755, Non-Activated Natural Killer Cell Engineering For Enhanced Cytotoxic Potential (1R44AI157181-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10139755. Licensed CC0.

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