沙特阿拉伯阿卜杜拉國王科技大學Christian Fr?kj?r-Jensen研究組發現，重編程piRNA通路可實現線蟲中多基因和跨代的基因沉默。相關論文于2022年2月3日在線發表于國際學術期刊《自然—方法學》。
研究人員發現，由染色體外轉基因表達的短的合成向導Piwi-interacting RNA（piRNA）（21個核苷酸的sg-piRNA）可以類似地重新編程內源性piRNA途徑，在線蟲雌雄同體的生殖系、精子和胚胎中進行基因特異性沉默。piRNA介導的干擾（'piRNAi'）比RNAi更有效，而且可以復用，生長素介導的piRNA特異性Argonaute PRG-1的降解能夠實現有條件的基因沉默。靶標特異性沉默的結果是信使RNA水平下降，二級小干擾RNA的擴增以及抑制性的染色質修飾。短的（300個堿基對）piRNAi轉基因從排列的寡核苷酸池中擴增出來，也能誘導沉默，有可能使piRNAi高度可擴展。
結果表明，piRNAi可以誘導兩個內源性基因（him-5和him-8）的跨代表觀遺傳學沉默。在靶標特異性sg-piRNA消失后，沉默會遺傳四到六代，而耗盡PRG-1則會導致基本永久性的表觀遺傳沉默。
據悉，單向導RNA可以將外源CRISPR-Cas蛋白靶向到獨特的DNA位置，使遺傳工具高效、特異且可擴展。
附：英文原文
Title: Reprogramming the piRNA pathway for multiplexed and transgenerational gene silencing in C. elegans
Author: Priyadarshini, Monika, Ni, Julie Zhouli, Vargas-Velazquez, Amhed M., Gu, Sam Guoping, Frkjr-Jensen, Christian
Issue&Volume: 2022-02-03
Abstract: Single-guide RNAs can target exogenous CRISPR–Cas proteins to unique DNA locations, enabling genetic tools that are efficient, specific and scalable. Here we show that short synthetic guide Piwi-interacting RNAs (piRNAs) (21-nucleotide sg-piRNAs) expressed from extrachromosomal transgenes can, analogously, reprogram the endogenous piRNA pathway for gene-specific silencing in the hermaphrodite germline, sperm and embryos of Caenorhabditis elegans. piRNA-mediated interference (‘piRNAi’) is more efficient than RNAi and can be multiplexed, and auxin-mediated degradation of the piRNA-specific Argonaute PRG-1 allows conditional gene silencing. Target-specific silencing results in decreased messenger RNA levels, amplification of secondary small interfering RNAs and repressive chromatin modifications. Short (300base pairs) piRNAi transgenes amplified from arrayed oligonucleotide pools also induce silencing, potentially making piRNAi highly scalable. We show that piRNAi can induce transgenerational epigenetic silencing of two endogenous genes (him-5 and him-8). Silencing is inherited for four to six generations after target-specific sg-piRNAs are lost, whereas depleting PRG-1 leads to essentially permanent epigenetic silencing.
DOI: 10.1038/s41592-021-01369-z
Source: nature.com/articles/s41592-021-01369-z
期刊信息
Nature Methods：《自然—方法學》，創刊于2004年。隸屬于施普林格·自然出版集團，最新IF：28.467
官方網址：https://www.nature.com/nmeth/
投稿鏈接：https://mts-nmeth.nature.com/cgi-bin/main.plex
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