Tech Session III: Gene Delivery and Gene Editing II
Lipid Nanoparticle Delivery for CRISPR Gene Editing in Primary Human T-cells
Wednesday, July 16, 2025
11:05 AM – 11:16 AM EDT
Introduction: Inborn errors of immunity (IEI) like hemophagocytic lymphohistiocytosis (HLH) may be treatable with T-cell gene correction. Lipid nanoparticles (LNPs) enable non-viral delivery of CRISPR machinery to a cell, but nuclear transport is a challenge. We previously showed truncated Cas9-target sequences (tCTS) enhance nuclear entry by allowing Cas9 ribonucleoprotein binding to HDR template (HDRT) and facilitating nuclear transport via a nuclear localization signal[2]. We identify LNPs that transfect primary human T cells and investigate adding tCTSs to improve large construct HDR knock-in (KI).
Learning Objectives:
At the completion of this activity, participants will know
Recognize the challenges of efficient DNA delivery to the nucleus for CRISPR editing.
Understand the impact of adding tCTS to HDRT for increasing nuclear delivery.
Describe the use of LNPs for translatable T-cell editing ex vivo for inborn errors of immunity.
