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Cells
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Research on CRISPR/Cas in Plants
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Research on CRISPR/Cas in Plants

A special issue of Cells (ISSN 2073-4409). This special issue belongs to the section "Plant, Algae and Fungi Cell Biology".

Deadline for manuscript submissions: closed (20 January 2022) | Viewed by 3247

Special Issue Editor

Dr. Randy Dinkins

E-Mail Website1 Website2
Guest Editor
USDA-ARS, Forage-Animal Production Research Unit, Lexington, KY 40546, USA
Interests: genetic improvement of crop plants utilizing molecular biology; tissue culture and transformation approaches

Special Issue Information

Dear Colleagues,

While discoveries using the CRISPR-Cas systems in plants have lagged behind animal systems, they are becoming an integral part of gene discovery, and future applications for crop breeding are expected to increase in the coming years. The current Special Issue aims to highlight successes, and obstacles encountered, utilizing the CRISPR-Cas system for gene discovery and genetic engineering especially in non-model crop species, where transformation techniques are more difficult and CRISPR-Cas targeting efficiency lower. The current Special Issue is calling for primary research articles or reviews that focus on current novel developments and applications that highlight the use of CRISPR-Cas technologies for gene discovery to plant breeding and advancement of agricultural products and applications.

Dr. Randy Dinkins
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Cells is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • CRISPR/Cas
  • Cas9
  • Cas12a
  • NHEJ
  • mutation
  • genome editing
  • gene knockout
  • gene knock-in
  • genetic engineering
  • agriculture

Published Papers (1 paper)

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Research

11 pages, 3153 KiB  
Article
LbCas12a-D156R Efficiently Edits LOB1 Effector Binding Elements to Generate Canker-Resistant Citrus Plants
by Hongge Jia, Yuanchun Wang, Hang Su, Xiaoen Huang and Nian Wang
Cells 2022, 11(3), 315; https://doi.org/10.3390/cells11030315 - 18 Jan 2022
Cited by 16 | Viewed by 2926
Abstract
Citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is an economically important disease in most citrus production regions worldwide. Xcc secretes a transcriptional activator like effector (TALE) PthA4 to bind to the effector binding elements (EBEs) in the promoter region of canker [...] Read more.
Citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is an economically important disease in most citrus production regions worldwide. Xcc secretes a transcriptional activator like effector (TALE) PthA4 to bind to the effector binding elements (EBEs) in the promoter region of canker susceptibility gene LOB1 to activate its expression, which in turn causes canker symptoms. Editing the EBE region with Cas9/gRNA has been used to generate canker resistant citrus plants. However, most of the EBE-edited lines generated contain indels of 1–2 bp, which has higher possibility to be overcome by PthA4 adaptation. The adaptation capacity of TALEs inversely correlates with the number of mismatches with the EBE. LbCas12a/crRNA is known to generate longer deletion than Cas9. In this study, we used a temperature-tolerant and more efficient LbCas12a variant (ttLbCas12a), harboring the single substitution D156R, to modify the EBE region of LOB1. We first constructed GFP-p1380N-ttLbCas12a:LOBP, which was shown to be functional via Xcc-facilitated agroinfiltration in Pummelo (Citrus maxima) leaves. Subsequently, we stably expressed ttLbCas12a:LOBP in Pummelo. Eight transgenic lines were generated, with seven lines showing 100% mutations of the EBE, among which one line is homozygous. The EBE-edited lines had the ttLbCas12a-mediated deletions of up to 10 bp. Importantly, the seven lines were canker resistant and no off-targets were detected. In summary, ttLbCas12a can be used to efficiently generate biallelic/homozygous citrus mutant lines with short deletions, thus providing a useful tool for the functional study and breeding of citrus. Full article
(This article belongs to the Special Issue Research on CRISPR/Cas in Plants)
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