2020 Jul 7;18(1):25. doi: 10.1186/s43141-020-00036-8. Transcriptional regulation with CRISPR-Cas9: principles, advances, and applications. Molecular mechanisms governing these complex root–soil–microbe interactions remain largely unknown, which hampers the development and use of reliable technologies like genome editing to engineer the components of rhizosphere for improved plant health and productivity. By using CRISPR-Cas9 targeting peanut AhNFR genes in hairy root transformation system, we validated the function of AhNFR5 genes in nodule formation in peanut. This study showed that CRISPR-Cas9 could be used in peanut hairy root transformation system for peanut functional genomic studies, specifically on the gene function in roots. in Plant Mol Biol 82(3):207-221, 2013). This editing technology has been in the limelight due to its simplicity and versatility compared to other previously known genome editing platforms. Epub 2019 Mar 15. The application of CRISPR/Cas9 in hairy roots to explore the functions of AhNFR1 and AhNFR5 genes during peanut nodulation. Find NCBI SARS-CoV-2 literature, sequence, and clinical content: https://www.ncbi.nlm.nih.gov/sars-cov-2/. Commons license, and indicate if changes were made. Taken together, the CRISPR/Cas9-mediated gene editing technology not only efficiently edited individual target gene but also showed satisfactory performances in multigene editing, which greatly promoted the development of genetic manipulations in filamentous fungi. PubMed Article First, we examined the transient genome editing activity of this system in tobacco protoplasts, insertion and deletion (indel) mutations were observed with frequencies of 16.2-20.3 % after transfecting guide RNA (gRNA) and the nuclease Cas9 in tobacco protoplasts. Latest Developed Strategies to Minimize the Off-Target Effects in CRISPR-Cas-Mediated Genome Editing. eCollection 2020. Thus, our results suggest that the synthetic transcriptional repressor (dCas9:SRDX) and activators (dCas9:EDLL and dCas9:TAD) can be used as endogenous transcription factors to repress or activate transcription of an endogenous genomic target. Abstract The relative ease, speed, and biological scope of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR … In vivo gene delivery mediated by non-viral vectors for cancer therapy. In this review, we discuss multiplexed CRISPR technologies and describe methods for the assembly, expression and processing of synthetic guide RNA arrays in vivo. While there is tremendous potential for CRISPR screens to map and interrogate gene regulatory networks at unprecedented speed and scale, their implementation in plants remains in its infancy. Applications that benefit from multiplexed CRISPR technologies, including cellular recorders, genetic circuits, biosensors, combinatorial genetic perturbations, large-scale genome engineering and the rewiring of metabolic pathways, are highlighted. All rights reserved. Display/Hide MLA, Chicago and APA citation formats. In order to streamline and facilitate rapid … This toolbox provides researchers with a protocol and reagents to quickly and efficiently assemble functional CRISPR/Cas9 transfer DNA constructs for monocots and dicots using Golden Gate and Gateway cloning methods. The protocol starts with an overview of the reagents and covers designing of gRNAs and assembly of components into a final T-DNA delivery molecule through Golden Gate cloning and Multisite Gateway LR recombination. Respiratory viral sepsis: epidemiology, pathophysiology, diagnosis and treatment. Combination of Cas9-Mediated Transcriptional Regulation With Synthetic Biology Here, we demonstrate that this strategy can be used for natural genes by CRISPR/Cas-mediated DSB induction. 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