小柯机器人

美国华盛顿大学Jay Shendure、Junhong Choi等研究人员合作开发出通过顺序基因组编辑的时间分辨的多符号分子记录器。2022年7月6日，《自然》杂志在线发表了这项成果。

据研究人员介绍，DNA自然很适合作为体内分子记录的数字媒介。然而，当代基于DNA的记忆装置在可同时记录的不同"符号"的数量方面受到限制，并且/或者无法捕捉事件发生的顺序。

研究人员报道了DNA打字机，一个用于体内分子记录的通用系统，它克服了这些和其他限制。对于DNA打字机，空白记录介质（"DNA磁带"）由部分CRISPR-Cas9目标位点的串联阵列组成，除了第一个位点外，所有位点都在其5′端被截断，因此没有活性。短暂的插入性编辑作为符号，记录了介导编辑的先导编辑向导RNA的身份，同时也将"类型向导"的位置沿着DNA胶带移动了一个单位，也就是顺序性的基因组编辑。

在这个DNA打字机的概念验证中，研究人员展示了对数千个符号、复杂事件历史和短文信息的记录和解码；评估了几十个正交磁带的性能；并构建了可能能够记录多达20个序列事件的'长磁带'。最后，研究人员将DNA打字机与单细胞RNA-seq结合起来，重建了一个由3257个细胞组成的单系，并发现对多拷贝DNA磁带的顺序编辑的泊松式积累可以在至少20代和25天的体外克隆扩增中保持。

附：英文原文

Title: A time-resolved, multi-symbol molecular recorder via sequential genome editing

Author: Choi, Junhong, Chen, Wei, Minkina, Anna, Chardon, Florence M., Suiter, Chase C., Regalado, Samuel G., Domcke, Silvia, Hamazaki, Nobuhiko, Lee, Choli, Martin, Beth, Daza, Riza M., Shendure, Jay

Issue&Volume: 2022-07-06

Abstract: DNA is naturally well suited to serve as a digital medium for in vivo molecular recording. However, contemporary DNA-based memory devices are constrained in terms of the number of distinct ‘symbols’ that can be concurrently recorded and/or by a failure to capture the order in which events occur1. Here we describe DNA Typewriter, a general system for in vivo molecular recording that overcomes these and other limitations. For DNA Typewriter, the blank recording medium (‘DNA Tape’) consists of a tandem array of partial CRISPR–Cas9 target sites, with all but the first site truncated at their 5′ends and therefore inactive. Short insertional edits serve as symbols that record the identity of the prime editing guide RNA2 mediating the edit while also shifting the position of the ‘type guide’ by one unit along the DNA Tape, that is, sequential genome editing. In this proof of concept of DNA Typewriter, we demonstrate recording and decoding of thousands of symbols, complex event histories and short text messages; evaluate the performance of dozens of orthogonal tapes; and construct ‘long tape’ potentially capable of recording as many as 20 serial events. Finally, we leverage DNA Typewriter in conjunction with single-cell RNA-seq to reconstruct a monophyletic lineage of 3,257 cells and find that the Poisson-like accumulation of sequential edits to multicopy DNA tape can be maintained across at least 20 generations and 25 days of in vitro clonal expansion.

DOI: 10.1038/s41586-022-04922-8

Source: https://www.nature.com/articles/s41586-022-04922-8

Nature：《自然》，创刊于1869年。隶属于施普林格·自然出版集团，最新IF：69.504
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