What does this research mean for the field?

The CDegSR system, utilizing a miniaturized and circularly permuted CRISPR-Cas13 degron-masking approach, enables high-contrast single-molecule imaging of endogenous RNA and reveals that nuclear crowding and translation machinery are the primary physical constraints on mRNA mobility. Novelty: ClaimNovelty.METHODOLOGICAL. Consensus alignment: ConsensusAlignment.NEUTRAL.

What question did this study set out to answer?

To engineer a compact, low-background CRISPR-Cas13 imaging system for tracking the spatiotemporal dynamics of unmodified endogenous RNA in living cells at single-molecule resolution.

September 2, 2026Advanced ScienceOpen Access

Single‐Molecule Imaging of Endogenous RNA Using a Miniaturized and Circularly Permuted CRISPR‐Cas13 Degron‐Masking System CDegSR

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Authors

SSShipeng ShaoSoochow University HZHongchen ZhangSouthwest University

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Implication

Experimental study demonstrates high-contrast single-molecule RNA imaging in living cells using an engineered CRISPR-Cas13 system, indicating nuclear crowding limits mRNA mobility.

Key Points

To engineer a compact, low-background CRISPR-Cas13 imaging system for tracking the spatiotemporal dynamics of unmodified endogenous RNA in living cells at single-molecule resolution.
Miniaturized dPspCas13b to its core recognition lobe and applied circular permutation to reposition the protein termini for target-dependent degron masking.
Designed the CDegSR effector to trigger rapid proteasomal degradation when free, while selectively stabilizing effectors bound to RNA targets.
Applied CDegSR to perform single-molecule tracking of endogenous POLR2A transcripts to evaluate intracellular mobility constraints.
Target-dependent degron masking selectively preserved target-bound complexes while degrading unbound effectors, substantially reducing background fluorescence.
Tracked individual endogenous POLR2A transcripts in real time, revealing that nuclear crowding and the translation machinery serve as the primary physical impediments to mRNA diffusion.

Cite This Study

Shao et al. (2026) studied this question.

synapsesocial.com/papers/6a97e2b1c562ede874ec6f5a https://doi.org/10.1002/advs.77533

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Implication

Experimental study demonstrates high-contrast single-molecule RNA imaging in living cells using an engineered CRISPR-Cas13 system, indicating nuclear crowding limits mRNA mobility.

Single‐Molecule Imaging of Endogenous RNA Using a Miniaturized and Circularly Permuted CRISPR‐Cas13 Degron‐Masking System CDegSR

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Implication

Key Points

Cite This Study

Single‐Molecule Imaging of Endogenous RNA Using a Miniaturized and Circularly Permuted CRISPR‐Cas13 Degron‐Masking System CDegSR

Discussion

Member takes

Implication

Key Points

Cite This Study

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