Single-cell DNA cytometry with magnetic- and fluorescence-activated bead sorting
Many clinically important cell populations are defined by intracellular DNA or RNA signatures, but isolating and profiling these cells remains difficult. Probe-based in situ hybridization strategies that are compatible with cytometry have difficulty resolving low-abundance transcripts, integrated provirus, or single-copy genomic mutations. In contrast, droplet digital PCR achieves single-molecule…
Researchers have developed a novel nucleic acid cytometry platform called magniFIND-seq. This innovative technology combines the high sensitivity of digital PCR with the accessibility of commercial magnetic-activated and fluorescence-activated sorting instruments.
In magniFIND-seq, individual cells are captured in agarose beads. Target sequences within these cells are then detected through digital PCR. After detection, the bead-bound amplicons are labeled with either magnetic or fluorescent probes. By utilizing controlled evaporation, the beads shrink from 55 to 20 micrometers in size. This size reduction enables scalable magnetic separation, fast single-bead FACS sorting, and their combined use for high-purity recovery of rare cell populations.
To demonstrate the capabilities of magniFIND-seq, researchers applied the platform to multiplexed single-copy detection and FACS-based isolation of specific targets. In one example, they successfully detected and isolated simian immunodeficiency virus proviral pol and env targets.
Additionally, magniFIND-seq was used to enrich chronic myeloid leukemia cells using the BCR::ABL1 target as a disease-defining marker. From this enrichment, single-cell transcriptomes were recovered, providing valuable insights into tyrosine kinase inhibitor-resistance programs.
With magniFIND-seq, researchers have extended the throughput and accessibility of nucleic acid cytometry. By engineering compatibility with commercial magnetic-activated and fluorescence-activated sorting instruments, this platform offers a more efficient and widely available solution for isolating and profiling clinically important cell populations based on their intracellular DNA or RNA signatures.
Written by urgent.news from bioRxiv's reporting — not their text. Machine-written — may contain errors; check the original before relying on it.
