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构建一组新型转座子载体,通过 CRISPR 干扰高效沉默蛋白和 lncRNA 基因。

Construction of a Set of Novel Transposon Vectors for Efficient Silencing of Protein and lncRNA Genes via CRISPR Interference.

机构信息

Department of Cell Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387, Kraków, Poland.

出版信息

Mol Biotechnol. 2023 Oct;65(10):1598-1607. doi: 10.1007/s12033-023-00675-5. Epub 2023 Jan 28.

Abstract

In recent years, CRISPR interference (CRISPRi) technology of gene silencing has emerged as a promising alternative to RNA interference (RNAi) surpassing the latter in terms of efficiency and accuracy. Here, we describe the construction of a set of transposon vectors suitable for constitutive or tetracycline (doxycycline)-inducible silencing of genes of interest via CRISPRi method and conferring three different antibiotic resistances, using vectors available via Addgene repository. We have analyzed the performance of the new vectors in the silencing of mouse Adam10 and human lncRNA, NORAD. The empty vector variants can be used to efficiently silence any genes of interest.

摘要

近年来,基因沉默的 CRISPR 干扰(CRISPRi)技术已经成为 RNA 干扰(RNAi)的一种有前途的替代方法,在效率和准确性方面超过了后者。在这里,我们描述了一组转座子载体的构建,这些载体适用于通过 CRISPRi 方法对感兴趣的基因进行组成型或四环素(强力霉素)诱导的沉默,并赋予三种不同的抗生素抗性,使用可通过 Addgene 存储库获得的载体。我们已经分析了新载体在沉默小鼠 Adam10 和人类 lncRNA NORAD 中的性能。空载体变体可用于有效地沉默任何感兴趣的基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/465f/10471651/d1c5c9ea582d/12033_2023_675_Fig1_HTML.jpg

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