a Laboratory of Experimental Virology, Department of Medical Microbiology , Amsterdam UMC, University of Amsterdam , Amsterdam , The Netherlands.
RNA Biol. 2018;15(12):1458-1467. doi: 10.1080/15476286.2018.1551703. Epub 2018 Nov 29.
The recently discovered clustered regularly interspaced short palindromic repeats (CRISPR)-Cpf1 system expands the genome editing toolbox. This system exhibits several distinct features compared to the widely used CRISPR-Cas9 system, but has reduced gene editing efficiency. To optimize the CRISPR-Cpf1 (Cas12a) system, we report the inclusion of self-cleaving ribozymes that facilitate processing of the crRNA transcript to produce the precise guide molecule. Insertion of the 3'-terminal HDV ribozyme boosted the gene editing activity of the CRISPR-Cpf1 system ranging from 1.1 to 5.2 fold. We also demonstrate that this design can enhance CRISPR-based gene activation. We thus generated an improved CRISPR-Cpf1 system for more efficient gene editing and gene regulation.
最近发现的成簇规律间隔短回文重复序列（CRISPR）-Cpf1 系统扩展了基因组编辑工具包。与广泛使用的 CRISPR-Cas9 系统相比，该系统具有几个明显的特点，但基因编辑效率降低。为了优化 CRISPR-Cpf1（Cas12a）系统，我们报告了包含自我切割核酶的方法，这有助于加工 crRNA 转录本以产生精确的指导分子。3'-末端 HDV 核酶的插入将 CRISPR-Cpf1 系统的基因编辑活性提高了 1.1 至 5.2 倍。我们还证明了这种设计可以增强基于 CRISPR 的基因激活。因此，我们生成了一种改进的 CRISPR-Cpf1 系统，用于更有效的基因编辑和基因调控。
