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Using intron-encoded cistronic transcripts coding and non-coding RNA are monitored.

Jonathan Dowell

Conventional gene reporters have a fundamental role in determining the physiological states of cells, but they can monitor gene expression while damaging proteins or significantly changing mature messenger RNA. Non-coding RNAs cannot currently be measured over multiple time points without having their nucleotide sequence changed, which can change their native function, half-life and localization. As a result, we created the minimally invasive transcriptional reporter intron-encoded scarless programmable extranuclear cistronic transcript (INSPECT), which is embedded within an intron of a gene of interest. After INSPECT's post-transcriptional excision, mature endogenous RNA without any sequence changes and another engineered transcript that leaves the taking control of the nuclear export system and translating it into a reporter or effector protein. We demonstrate its application in tracking the transcriptional dynamics of the long non-coding RNA (lncRNA) NEAT1 during CRISPR interference-mediated perturbation and monitoring interleukin-2 (IL2) after T cell activation. The technique known as INSPECT allows for the monitoring of gene transcription without affecting the target's mature messenger or lncRNA.