MIBE Online Seminar: Intron-Encoded Cistronic Transcripts for Minimally-Invasive Monitoring of (Non-)Coding RNAs

Titel: Intron-Encoded Cistronic Transcripts for Minimally-Invasive Monitoring of (Non-)Coding RNAs.

Sprecher: Niklas Armbrust, HELENA doctoral candidate (Westmeyer Group)

Datum und Uhrzeit: Dienstag, 08. Februar 2022, 13:00 Uhr

Abstract: Despite their fundamental role for assessing (patho)physiological cell states, conventional gene reporters can follow gene expression but leave scars on the proteins or substantially alter the mature mRNA. Multi-time-point measurements of non-coding RNAs are currently impossible without modifying their nucleotide sequence, which can alter their native function, half-life, and localization. We thus developed INSPECT, Intron-encoded Scarless Programmable Extranuclear Cistronic Transcripts, as minimally-invasive transcriptional reporters embedded within an intron of a gene of interest. Post-transcriptional excision of INSPECT results in unmodified mRNA and a synthetic transcript that leaves the nucleus by hijacking the nuclear export machinery for subsequent translation into a reporter or effector protein. We showcase monitoring interleukin-2 (IL2) after T-cell activation and tracking transcriptional dynamics of the long non-coding RNA (lncRNA) NEAT1 during CRISPRi-mediated perturbation. INSPECT is the first method for monitoring gene transcription without altering the target of interest at either the RNA or protein level.