ZFP36L2 orchestrates stress-adaptive plasticity in intestinal regeneration and colorectal cancer metastasis (SLAMseq)

Organoids expressing a doxycycline-inducible short hairpin RNA targeting ZFP36L2 (shZFP36L2) or a non-targeting control (shCtrl) were metabolically labeled with 500 μM 4-thiouridine (4sU) for 6 hours, followed by a uridine chase.

Total RNA was harvested in triplicate at t=0, 2, 5, and 12 hours post-chase, along with matched unlabeled control samples for background T>C correction. RNA was subjected to iodoacetamide alkylation and libraries were prepared using the QuantSeq 3′ mRNA-seq V2 Library Prep Kit FWD (Lexogen). T>C conversion rates were used to quantify labeled RNA fractions and fit single-exponential decay models to estimate per-gene mRNA half-lives.