S. Patange, S. Maragh
U.S. National Institute of Standards and Technology,
United States
Keywords: single-cell, FluidFM, genome editing, gene editing, NIST
Summary:
Genome editing is a rapidly evolving biotechnology with the potential to transform many sectors of industry including agriculture, biomanufacturing, and medicine. Genome editing consists of a portfolio of biomolecular techniques that introduce site-specific modifications to the genetic code of living cells. In order to introduce genome editing biomolecules into cells, one must use a reagent delivery method that could either manipulate cells in bulk or at the single-cell level. One emerging technology is Fluid Force Microscopy (1), which uses AFM principles to control a microfluidic probe for precise single-cell manipulation and controlled reagent delivery. In this talk, we present our results using a commercial instrument based on this technology, the ‘FluidFM OMNIUM’ (Cytosurge AG). We present our data on the instrument’s measurement capabilities such as spatial recall, fluorescence detection, and microfluidic flow analysis— as well as its potential for delivering genome editing reagents into human cells. This work builds upon guidance provided in our recent publication (2), in which we summarize technologies related to research applications where it is advantageous to have controlled dose delivery of a reagent, and/or high-resolution data on the distribution of genomic and phenotypic outcomes of cells that were individually manipulated at the single-cell level. References: (1) Meister et al., Nano Lett. 2009: DOI: 10.1021/nl901384x (2) Patange and Maragh, Biochemistry 2022, DOI: 10.1021/acs.biochem.2c00431