Bi Youkun, Gu Lei, Wang Jun, Chang Yan, Jin Meng, Mao Yang, Wang Huiwen, Ji Guangju
Key Laboratory of Interdisciplinary Research, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
University of Chinese Academy of Sciences, Beijing, China.
CRISPR J. 2021 Jun;4(3):381-391. doi: 10.1089/crispr.2020.0110. Epub 2021 Jun 2.
Recombinant adenoviruses have broad applications for gene delivery and expression. Furthermore, the adenovirus packaging system facilitates the expression of RNA-guided CRISPR/Cas9 nuclease complexes. In this study, we developed a novel system, named AdBlue, for the construction of recombinant adenoviruses using an enzymatic assembly strategy. This system could significantly reduce the time and labor required to generate adenoviral vectors. When applied to CRISPR/Cas9 design, it simplifies the preparation of recombinant adenoviruses carrying nuclease complexes and can induce high levels of site-specific mutagenesis. Our system has outstanding advantages for adenovirus preparation and could be a useful molecular engineering tool for gene delivery and editing.
重组腺病毒在基因递送和表达方面有广泛应用。此外，腺病毒包装系统有助于RNA引导的CRISPR/Cas9核酸酶复合物的表达。在本研究中，我们开发了一种名为AdBlue的新型系统，用于采用酶促组装策略构建重组腺病毒。该系统可显著减少产生腺病毒载体所需的时间和人力。当应用于CRISPR/Cas9设计时，它简化了携带核酸酶复合物的重组腺病毒的制备，并可诱导高水平的位点特异性诱变。我们的系统在腺病毒制备方面具有突出优势，可能是一种用于基因递送和编辑的有用分子工程工具。
