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An imaging tool follows RNA through living plant cells with less background

A peer-reviewed plant experiment developed a fluorescent imaging system that clears unbound probes while keeping signals attached to RNA. The team followed a flowering-related RNA at channels connecting cells and in grafted tissues. Tests in two model plants reduced background and improved contrast, while the large imaging complex and limited reporter configuration constrain current use.

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A green leaf on a glass slide beneath a microscope objective, with a potted plant and leaf samples behind it in a laboratory.

Free probes are cleared while bound signals remain

A peer-reviewed experiment developed a CRISPR-based fluorescent tool to follow RNA, molecules that carry genetic messages, in living plant cells.[1], [2]

The system preserves signals attached to the target while reducing background from probes that remain unbound. A plant cell's large central vacuole squeezes cytoplasm into a narrow outer layer, where free probes can accumulate and obscure the image.[1]

The researchers combined dCas13X, a protein that recognizes RNA, with SunTag, a structure that amplifies the fluorescent signal. A cPIL degradation signal helps the cell's protein-disposal machinery clear unbound probes. The experiments used Nicotiana benthamiana and Arabidopsis thaliana, two plant species used as laboratory models.[1]

Flowering-related RNA crosses cell channels

The system visualized RNA for FT, a flowering signal, near plasmodesmata, the channels linking plant cells. RNA first remained close to a channel before crossing the boundary. Grafting experiments also detected FT signals in recipient tissues and separately confirmed full-length RNA. Transport declined in plants lacking GRP7 protein; a role in a broader RNA-protein complex remains possible, without establishing an FT-specific direct interaction.[1]

Contrast improves in the tested configuration

In the compared configurations, background decreased 6.71-fold and the signal-to-noise ratio rose 6.67-fold. The team detected no measurable change in the tested targets' RNA abundance, protein products or RNA decay. The imaging complex remains relatively large, and the degradation module was validated only with green fluorescence in a single-color setting. Those results apply to the tested plants and targets rather than establishing the same performance across all species.[1]

References

  1. News sourcePLOS BiologyA binding-activated CRISPR/dCas13X imaging system enables precise RNA tracking in plants↩1↩2↩3↩4↩5
  2. News sourceCiencias.uyCRISPR imaging follows RNA in living plants with less background↩
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