Harnessing CRISPR-Cas for oomycete genome editing

Oomycetes are a group of microorganisms that include pathogens responsible for devastating diseases in plants and animals worldwide. Despite their importance, the development of genome editing techniques for oomycetes has progressed more slowly than for model microorganisms. Here, we review recent breakthroughs in clustered regularly interspaced short palindromic repeats (CRISPR)-Cas technologies that are expanding the genome editing toolbox for oomycetes – from the original Cas9 study to Cas12a editing, ribonucleoprotein (RNP) delivery, and complementation. We also discuss some of the challenges to applying CRISPR-Cas in oomycetes and potential ways to overcome them. Advances in CRISPR-Cas technologies are being used to illuminate the biology of oomycetes, which ultimately can guide the development of tools for managing oomycete diseases.     

2017基因组编辑专刊序言

基因组编辑技术,作为一项生物医学领域的革新技术,已经在动物、植物和微生物基因组改造中得到了广泛的应用。以CRISPR/Cas9为主导的基因组编辑技术掀起了基因组编辑的浪潮,在功能基因组学、遗传改良育种、遗传病治疗等研究中展示出其极大的价值与潜力。本专刊报道了基因组编辑技术的总体状况、在相关领域的基础与应用研究、该技术当前存在的优缺点以及未来展望等。     

DNA-guided genome editing tool

正Technologies for targeted modification of eukaryotic genomes are of great value for elucidating and manipulating gene functions in fundamental and applied biological research.In the past two decades,genome editing tools have evolved quickly from random DNA mutagenesis-inducing chemicals or ionizing radiation to programmable sequencespecific nucleases(SSNs)with the CRISPR/Cas technology as the most significant breakthrough(Gaj et al.,2013).     

Current and future editing reagent delivery systems for plant genome editing

Many genome editing tools have been developed and new ones are anticipated; some have been extensively applied in plant genetics, biotechnology and breeding, especially the CRISPR/Cas9 system. These technologies have opened up a new era for crop improvement due to their precise editing of user-specified sequences related to agronomic traits. In this review, we will focus on an update of recent developments in the methodologies of editing reagent delivery, and consider the pros and cons of current delivery systems. Finally, we will reflect on possible future directions.     

CRISPR-derived genome editing technologies for metabolic engineering

In metabolic engineering, genome editing tools make it much easier to discover and evaluate relevant genes and pathways and construct strains. Clustered regularly interspaced palindromic repeats (CRISPR)-associated (Cas) systems now have become the first choice for genome engineering in many organisms includingindustrially relevant ones. Targeted DNA cleavage by CRISPR-Cas provides variousgenome engineering modes such as indels, replacements, large deletions, knock-in and chromosomal rearrangements, while host-dependent differences in repair pathways need to be considered. The versatility of the CRISPR system has given rise to derivative technologies that complement nuclease-based editing, which causes cytotoxicity especially in microorganisms. Deaminase-mediated base editing installs targeted point mutations with much less toxicity. CRISPRi and CRISPRa can temporarily control gene expression without changing the genomic sequence. Multiplex, combinatorial and large scale editing are made possible by streamlined design and construction of gRNA libraries to further accelerates comprehensive discovery, evaluation and building of metabolic pathways. This review summarizes the technical basis and recent advances in CRISPR-related genome editing tools applied for metabolic engineering purposes, with representative examples of industrially relevant eukaryotic and prokaryotic organisms.     

植物基因组编辑新工具——引导编辑技术

基于CRISPR/Cas9系统的引导编辑(prime editing,PE)技术作为一种新兴的基因组编辑技术,能在不产生双链断裂的情况下实现所有12种单碱基替换和小片段DNA的缺失或插入.引导编辑技术已经在多种植物中成功应用并将在植物精准育种中发挥重要作用.虽然植物引导编辑(plant prime editing,PP...     

Barriers to genome editing with CRISPR in bacteria

Journal of Industrial Microbiology & Biotechnology - Genome editing is essential for probing genotype–phenotype relationships and for enhancing chemical production and phenotypic...     

Biocommunication and natural genome editing

The biocommunicative approach investigates communication processes within and among cells, tissues, organs and organisms as sign-mediated interactions, and nucleotide sequences as code, i.e. language-like text, which follows in parallel three kinds of rules: combinatorial (syntactic), context-sensitive (pragmatic), and content-specific (semantic). Natural genome editing from a biocommunicative perspective is competent agent-driven generation and integration of meaningful nucleotide sequences into pre-existing genomic content arrangements and the ability to (re-)combine and (re-)regulate them according to context-dependent (i.e. adaptational) purposes of the host organism.     

New cytosine base editor for plant genome editing

正A large number of beneficial agronomic traits in crops are associated with single nucleotide polymorphisms (SNPs) or point mutations (Jiao et al., 2010; Li et al., 2017; Ma et al.,2015). In the past, site-specific point mutations in a target gene can only be achieved through the CRISPR/Cas9-mediated gene replacement via the homology-directed repair(HDR). However, the intrinsically low HDR activity in plant cells and the lack of efficient way to supply abundant HDR templates in plant nucleus have greatly limited the success     

DNA-free genome editing methods for targeted crop improvement

Evolution of the next-generation clustered, regularly interspaced, short palindromic repeat/Cas9 (CRISPR/Cas9) genome editing tools, ribonucleoprotein (RNA)-guided endonuclease (RGEN) RNPs, is paving the way for developing DNA-free genetically edited crop plants. In this review, I discuss the various methods of RGEN RNPs tool delivery into plant cells and their limitations to adopt this technology to numerous crop plants. Furthermore, focus is given on the importance of developing DNA-free genome edited crop plants, including perennial crop plants. The possible regulation on the DNA-free, next-generation genome-edited crop plants is also highlighted.