# Acoustic microvortices instrumentation for dosage controlled, high efficiency cell engineering

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA-IRVINE · 2024 · $295,827

## Abstract

Contact PD/PI: LEE, ABRAHAM
Acoustic microvortices instrumentation for dosage controlled, high efficiency cell engineering
Abstract - In years 2018 and 2020, Nobel Prizes were awarded to pioneers in cancer immunotherapy
and the gene editing CRISPR-Cas9 method. This has ushered in a new era of cell engineering that
demands technological innovations to produce genetic-modified and reprogrammed immune cells (e.g.,
T cells). CAR T cell immunotherapy is one type of cell therapy that has already achieved success in
the clinic for treating cancer. The holy grail is to produce universal CAR T cells, genetically modified
and engineered allogeneic T cells derived from healthy donors that avoid immunologic rejection. This
requires gene engineering techniques such as CRISPR-Cas9 that can deliver multiplex gene insertions
and knock-outs with controlled dosage to enhance viability and efficacy of the engineered cells.
Although viral vectors are the method of choice for cell engineering, there are major concerns over their
safety as well as the complex, costly preparation process. Nonviral transfection methods are
alternatives to viral vectors that avoid the deleterious byproducts such as immune-mediated toxicity
and oncogenic transgene concerns. Here we introduce the Acoustic-Electric Shear Orbiting Poration
(AESOP) platform that addresses several of the known challenges for nonviral transfection techniques,
including cell viability and dosage-controlled intracellular delivery while achieving relatively high
throughput. AESOP is based on a microfluidic platform termed “lateral cavity acoustic transducers
(LCATs)” that can form an array of microvortices activated by acoustic energy. The main innovation
of the AESOP platform is the trapping of suspended populations of cells in tunable 3-D
microvortices and incorporating a two-step membrane disruption strategy to precisely
permeabilize cell membranes. By trapping cells to tumble in whirlpool-like microvortices, this platform
optimizes the delivery of intended cargo sizes with uniform poration of the cell membranes via
mechanical shear followed by the modulated enlargement of these nanopores via electric field. Using
AESOP, we will focus on technology development for producing the universal CAR T cells. By
controlling dosage, a decreased amount of CRISPR reagents in cells could reduce off-target effects.
Furthermore, the uniform delivery enables the delivery of large cargo sizes necessary for
immunotherapy-related gene transfection and gene editing. This 4-year project has four specific
aims: 1- Generate acoustic microstreaming vortices for uniform cell membrane nanopores and
uniform local mixing; 2- Demonstrate uniform electric field enlargement of nanopores for cargo
delivery; 3- Design prototype AESOP instrumentation and 4- Quantitative benchmark of AESOP
for intracellular delivery of large size cargos with dosage control for the development of
universal CAR T cells.

## Key facts

- **NIH application ID:** 10874449
- **Project number:** 5R01GM145987-03
- **Recipient organization:** UNIVERSITY OF CALIFORNIA-IRVINE
- **Principal Investigator:** Abraham P Lee
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $295,827
- **Award type:** 5
- **Project period:** 2022-06-01 → 2026-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10874449

## Citation

> US National Institutes of Health, RePORTER application 10874449, Acoustic microvortices instrumentation for dosage controlled, high efficiency cell engineering (5R01GM145987-03). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10874449. Licensed CC0.

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