加州理工学院王开航小组近日取得一项新成果。经过不懈努力，他们研究出利用DNA三向连接构建复杂多样的DNA序列。这一研究成果发表在2026年1月21日出版的国际学术期刊《自然》上。

为了从根本上解决这一挑战，课题组在这里展示了一种名为响尾蛇的新DNA组装技术的发展，该技术将指导组装的信息从最终组装序列中分离出来，这些序列以DNA三方连接为主题。该研究团队展示了Sidewinder技术的变革性质：高度自动化和准确地构建了40个片段的多片段组装，高GC含量和高重复的复杂DNA序列，在同一反应中平行组装多个不同基因，以及在整个基因长度上具有大量不同位置的组合文库，以实现442368个变体文库的高覆盖。该技术可实现高保真DNA组装，在三向连接处的误连接率约为百万分之一。

研究人员表示，从头构建全新的合成DNA序列的能力对工程和生物学研究至关重要。然而，产生长而复杂的合成DNA序列和文库的能力目前落后于测序和编辑DNA的能力。所有现有的DNA组装技术都依赖于在最终结构中发现的DNA序列信息来指导DNA分子之间的组装。这种模式的结果是，这些序列不可能在不影响最终序列的情况下被广泛优化。

附：英文原文

Title: Construction of complex and diverse DNA sequences using DNA three-way junctions

Author: Robinson, Noah Evan, Zhang, Weilin, Ghosh, Rajesh, Gerber, Bryan, Zhang, Hanqiao, Sanfiorenzo, Charles, Wang, Sixiang, Di Carlo, Dino, Wang, Kaihang

Issue&Volume: 2026-01-21

Abstract: The ability to construct entirely new synthetic DNA sequences de novo is essential to engineering and studying biology. However, the ability to produce long complex synthetic DNA sequences and libraries currently lags behind the ability to sequence and edit DNA1,2. All existing DNA-assembly technologies rely on DNA sequence information found within the final construct to direct assembly between DNA molecules3,4,5,6,7,8,9,10,11. As a result of this paradigm, these sequences cannot be extensively optimized specifically for assembly without affecting the final sequence. To fundamentally address this challenge, here we show the development of a new DNA assembly technique named Sidewinder that separates the information that guides assembly from the final assembled sequence using DNA three-way junctions. We demonstrate the transformative nature of the Sidewinder technique with highly robust and accurate construction of a 40-piece multifragment assembly, complex DNA sequences of both high GC content and high repeats, parallel assembly of multiple distinct genes in the same reaction and a combinatorial library with a large number of diversified positions across the entire length of the gene for high coverage of a library of 442,368 variants. This technology enables high-fidelity DNA assembly with a misconnection rate at the three-way junction of approximately 1 in 1,000,000.

DOI: 10.1038/s41586-025-10006-0

Source: https://www.nature.com/articles/s41586-025-10006-0