德国慕尼黑亥姆霍兹中心Gil Gregor Westmeyer团队近期取得重要工作进展，他们研究开发了内含子编码的顺反子转录本，用于编码和非编码RNA的微创监测。相关论文2022年11月7日在线发表于《自然-细胞生物学》杂志上。

因此，研究人员开发了内含子编码的无疤痕可编程核外顺反子转录物（INSPECT），作为嵌入感兴趣基因内含子内的微创转录报告物。INSPECT的转录后切除导致成熟的内源性RNA没有序列改变，并通过劫持核输出机制而离开细胞核，随后翻译为报告蛋白或效应蛋白。研究人员展示了其在监测T细胞活化后的白细胞介素-2（IL2）以及在CRISPR干扰介导的扰动期间，跟踪长链非编码RNA（lncRNA）NEAT1的转录动力学中的应用。INSPECT是一种监测基因转录动态而不改变靶标位点成熟lncRNA或mRNA的方法。

据介绍，尽管传统的基因报告子在评估（病理）生理细胞状态中起着基本作用，可以跟踪基因的表达，但会在蛋白质上留下疤痕或显著改变成熟的mRNA。非编码RNA的多时间点测量需要修改其核苷酸序列，这会改变其天然功能、半衰期和定位。

附：英文原文

Title: Intron-encoded cistronic transcripts for minimally invasive monitoring of coding and non-coding RNAs

Author: Truong, Dong-Jiunn Jeffery, Armbrust, Niklas, Geilenkeuser, Julian, Lederer, Eva-Maria, Santl, Tobias Heinrich, Beyer, Maren, Ittermann, Sebastian, Steinmal, Emily, Dyka, Mariya, Raffl, Gerald, Phlairaharn, Teeradon, Greisle, Tobias, ivani, Milica, Grosch, Markus, Drukker, Micha, Westmeyer, Gil Gregor

Issue&Volume: 2022-11-07

Abstract: Despite their fundamental role in assessing (patho)physiological cell states, conventional gene reporters can follow gene expression but leave scars on the proteins or substantially alter the mature messenger RNA. 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. Thus, we developed the intron-encoded scarless programmable extranuclear cistronic transcript (INSPECT) as a minimally invasive transcriptional reporter embedded within an intron of a gene of interest. Post-transcriptional excision of INSPECT results in the mature endogenous RNA without sequence alterations and an additional engineered transcript that leaves the nucleus by hijacking the nuclear export machinery for subsequent translation into a reporter or effector protein. We showcase its use in monitoring interleukin-2 (IL2) after T cell activation and tracking the transcriptional dynamics of the long non-coding RNA (lncRNA) NEAT1 during CRISPR interference-mediated perturbation. INSPECT is a method for monitoring gene transcription without altering the mature lncRNA or messenger RNA of the target of interest.

DOI: 10.1038/s41556-022-00998-6

Source: https://www.nature.com/articles/s41556-022-00998-6