Advertisement

Topics

Cas9-NG greatly expands the targeting scope of genome-editing toolkit by recognizing NG and other atypical PAMs in rice.

08:00 EDT 27th March 2019 | BioPortfolio

Summary of "Cas9-NG greatly expands the targeting scope of genome-editing toolkit by recognizing NG and other atypical PAMs in rice."

The CRISPR technologies enabling precise genome manipulation are valuable for the gene function study and molecular crop breeding. However, the requirement of a protospacer adjacent motif (PAM), such as NGG, TTN, etc., for Cas protein recognition restricts the targetable genomic loci in applications. Very lately, Cas9-NG which recognizes a minimal NG PAM was reported to expand the targeting space in genome editing in human cells, but little is known about its applications in plants. Here, we evaluated the nuclease activity of Cas9-NG toward various NGN PAMs by targeting endogenous genes in transgenic rice, and revealed that Cas9-NG edits all NGG, NGA, NGT and NGC sites with impaired activities, and gene-edited plants were dominated by mono-allelic mutations. The Cas9-NG-engineered base editors were further developed and used to generate OsBZR1 gain-of-function plants, which other available Cas9-engineered base editors are not able to. Moreover, Cas9-NG-based transcriptional activator efficiently up-regulated the expression of endogenous target genes in rice. In addition, we discovered that Cas9-NG recognizes NAC, NTG, NTT, NCG apart from NG PAM. All these findings show that Cas9-NG greatly expands the targeting scope of genome-editing tools and imply great potentials for targeted genome editing, base editing and genome regulation in plants.

Affiliation

Journal Details

This article was published in the following journal.

Name: Molecular plant
ISSN: 1752-9867
Pages:

Links

DeepDyve research library

PubMed Articles [9969 Associated PubMed Articles listed on BioPortfolio]

Improving plant genome editing with high-fidelity xCas9 and non-canonical PAM-targeting Cas9-NG.

Engineered SpCas9 variants, namely xCas9 and Cas9-NG, have shown promising potential on improving targeting specificity and broadening the targeting range. In this study, we evaluated these Cas9 varia...

Developing a highly efficient and wildly adaptive CRISPR-SaCas9 toolset for plant genome editing.

The CRISPR- Streptococcus pyogenes Cas9 (SpCas9) system offers a rapid, simple and flexible genome editing approach. However, the targeting scope of the SpCas9 system is limited by the canonical NGG P...

A Single Transcript CRISPR-Cas9 System for Multiplex Genome Editing in Plants.

The CRISPR-Cas9 system has been widely adopted in genome editing. By changing the 20 bp guide sequence, it can easily edit any sequence adjacent to a protospacer adjacent motif (PAM) in a genome. Mul...

Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha.

Marchantia polymorpha is one of the model species of basal land plants. Although CRISPR/Cas9-based genome editing has already been demonstrated for this plant, the efficiency was too low to apply to f...

A new approach to Cas9-based genome editing in Aspergillus niger that is precise, efficient and selectable.

Aspergillus niger and other filamentous fungi are widely used in industry, but efficient genetic engineering of these hosts remains nascent. For example, while molecular genetic tools have been develo...

Clinical Trials [1764 Associated Clinical Trials listed on BioPortfolio]

Ascending Dose Study of Genome Editing by the Zinc Finger Protein (ZFP) Therapeutic SB-318 in Subjects With MPS I

The purpose of the study is to evaluate the safety, tolerability and effect on leukocyte and plasma iduronidase (IDUA) enzyme activity of ascending doses of SB-318. SB-318 is an intravenou...

Ascending Dose Study of Genome Editing by the Zinc Finger Nuclease (ZFN) Therapeutic SB-913 in Subjects With MPS II

The purpose of the study is to evaluate the safety, tolerability and effect on leukocyte and plasma Iduronate 2-Sulfatase (IDS) enzyme activity of ascending doses of SB-913. SB-913 is an i...

Cell Therapy for High Risk T-Cell Malignancies Using CD7-Specific CAR Expressed On Autologous T Cells

Patients eligible for this study have a type of blood cancer called T-cell leukemia or lymphoma (lymph gland cancer). The body has different ways of fighting infection and disease. This s...

The Use of a Language Toolkit for Toddlers

To investigate whether young children with isolated expressive language delay benefit from early intervention with a simple language toolkit and brief instructions provided to their caregi...

Exploiting Epigenome Editing in Kabuki Syndrome: a New Route Towards Gene Therapy for Rare Genetic Disorders

Starting from isolating primary cells from affected patients, an in vitro disease model system for KS will be developed. Using alternative strategies to obtain patient-derived mesenchymal ...

Medical and Biotech [MESH] Definitions

Genetic engineering techniques that involve DNA REPAIR mechanisms for incorporating site-specific modifications into a cell's genome.

The integration of exogenous DNA into the genome of an organism at sites where its expression can be suitably controlled. This integration occurs as a result of homologous recombination.

An APOBEC deaminase catalytic subunit of the apolipoprotein B (APOB) MESSENGER RNA (mRNA) editing enzyme complex that is involved in post-transcriptional editing of a CAA codon for GLYCINE to a UAA STOP CODON in the ApoB mRNA. It also functions in CGA (ARGININE) to UGA STOP CODON editing of NEUROFIBROMIN 1 mRNA and EPIGENETIC PROCESSES.

Instruments for the visual examination of interior structures related to NEUROSURGICAL PROCEDURES. There are two broad categories of neuroendoscopes, the rigid scope and the flexible scope, for various types of procedures.

A process that changes the nucleotide sequence of mRNA from that of the DNA template encoding it. Some major classes of RNA editing are as follows: 1, the conversion of cytosine to uracil in mRNA; 2, the addition of variable number of guanines at pre-determined sites; and 3, the addition and deletion of uracils, templated by guide-RNAs (RNA, GUIDE).

Advertisement
Quick Search
Advertisement
Advertisement

 


DeepDyve research library

Relevant Topics

Bioinformatics
Bioinformatics is the application of computer software and hardware to the management of biological data to create useful information. Computers are used to gather, store, analyze and integrate biological and genetic information which can then be applied...

Nutrition
Within medicine, nutrition (the study of food and the effect of its components on the body) has many different roles. Appropriate nutrition can help prevent certain diseases, or treat others. In critically ill patients, artificial feeding by tubes need t...


Searches Linking to this Article