The Shanghai Academy of Health Sciences uses CRISPR/Cpf1 system to achieve multi-gene fixed-point editing of rice.

On March 19, the journal Molecular Plant published online a research paper entitled Multiplex Gene Editing in Rice Using THE CRISPR-Cpf1 System by Zhu Health Research Group of the Center for Plant Adversity Biology, Shanghai Institute of Life Sciences, Chinese Academy of Sciences.
this work in rice using CRISPR/Cpf1 system to achieve multi-gene site editing, the efficiency of 40-75%, and the system in the vector construction than CRISPR/Cas9 system is easier.
this work provides a simple and efficient new tool for rice multi-gene spot editing.
THE CRISPR/Cas9 system has been widely used in gene fixed-point editing, however, the CRISPR/Cas9 system still has some limitations.
in crispr/Cas9 systems, CRISPR transcription is required to form two small RNA molecules, RNA CRISPR RNA (CrRNA) and trans-acting CrRNA (trans-activated crRNA, tracrRNA), which are paired to form complexes in parts of the RNA molecule region.
, the complex guides Cas proteins with nonse specific nuclease activity to cut DNA sequences that match crRNA.
, the engineered CRISPR/Cas9 system also needs to fuse CrRNA with tracerRNA to form a single chimeric RNA (chiRNA) and need the help of the host RNA enzyme to function.
same time, when using the system for a single multi-gene fixed-point editing, the carrier system is complex, and each fixed-point editing bit requires a separate starter and termination series.
In recent years, scientists have discovered and modified the CRISPR from Prevotella and Francisella 1 systems, which have shown that the system overcomes the limitations of CRISPR/Cas9, does not require the assistance of tracerRNA, and that the CF1 protein, which combines the functions of DNA shearing enzymes and RNA trimming enzymes, not only targets the cutting of DNA double strands, but also processes the corresponding immature CrRNA (pre-crRNA) into mature CrRNA.
health research team used Francisella novicida Cpf1 (FnCpf1) and Lachnospiraceae Bacterium ND2006 Cpf1 (LbCpf1) to test rice for unit and multiple point gene knocks, and studies have shown that the two Cpf1s require only one A very short direct repeat sequence of 20-21bp (direct repeats, DR) plus a 22-24bp target identification sequence (guide) enables single gene knock-off and, more importantly, multiple DR-guide units are directly connected in series, requiring only one initiation driver to achieve simple and efficient multi-gene knock-out.
the study used a crRNA short array of four DR-guide units to edit four genes in the rice RLK and CYP81A families, each with a knock-out efficiency of 40-75%.
the system is simple and efficient to achieve multi-gene fixed-point editing in rice, expand the application of CRISPR system in plants, and provide a new weapon for rice genome fixed-point editing.
the work has been supported by funding from the Chinese Academy of Sciences.
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