Construct design for CRISPR/Cas-based genome editing in plants

Md Mahmudul Hassan¹, Yingxiao Zhang², Guoliang Yuan³, Kuntal De⁴, Jin-Gui Chen³, Wellington Muchero³, Gerald A Tuskan³, Yiping Qi⁵, Xiaohan Yang⁶

Affiliations

¹ Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA; The Center for Bioenergy Innovation, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA; Department of Genetics and Plant Breeding, Patuakhali Science and Technology University, Dumki, Patuakhali-8602, Bangladesh.
² Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD 20742, USA.
³ Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA; The Center for Bioenergy Innovation, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA.
⁴ Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA.
⁵ Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD 20742, USA; Institute for Bioscience and Biotechnology Research, University of Maryland, Rockville, MD 20850, USA. Electronic address: yiping@umd.edu.
⁶ Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA; The Center for Bioenergy Innovation, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA. Electronic address: yangx@ornl.gov.

PMID: 34340931
DOI: 10.1016/j.tplants.2021.06.015

Free article

Review

Construct design for CRISPR/Cas-based genome editing in plants

Md Mahmudul Hassan et al. Trends Plant Sci. 2021 Nov.

Free article

. 2021 Nov;26(11):1133-1152.

doi: 10.1016/j.tplants.2021.06.015. Epub 2021 Jul 31.

Authors

Md Mahmudul Hassan¹, Yingxiao Zhang², Guoliang Yuan³, Kuntal De⁴, Jin-Gui Chen³, Wellington Muchero³, Gerald A Tuskan³, Yiping Qi⁵, Xiaohan Yang⁶

Affiliations

¹ Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA; The Center for Bioenergy Innovation, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA; Department of Genetics and Plant Breeding, Patuakhali Science and Technology University, Dumki, Patuakhali-8602, Bangladesh.
² Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD 20742, USA.
³ Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA; The Center for Bioenergy Innovation, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA.
⁴ Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA.
⁵ Department of Plant Science and Landscape Architecture, University of Maryland, College Park, MD 20742, USA; Institute for Bioscience and Biotechnology Research, University of Maryland, Rockville, MD 20850, USA. Electronic address: yiping@umd.edu.
⁶ Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA; The Center for Bioenergy Innovation, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA. Electronic address: yangx@ornl.gov.

PMID: 34340931
DOI: 10.1016/j.tplants.2021.06.015

Abstract

CRISPR construct design is a key step in the practice of genome editing, which includes identification of appropriate Cas proteins, design and selection of guide RNAs (gRNAs), and selection of regulatory elements to express gRNAs and Cas proteins. Here, we review the choices of CRISPR-based genome editors suited for different needs in plant genome editing applications. We consider the technical aspects of gRNA design and the associated computational tools. We also discuss strategies for the design of multiplex CRISPR constructs for high-throughput manipulation of complex biological processes or polygenic traits. We provide recommendations for different elements of CRISPR constructs and discuss the remaining challenges of CRISPR construct optimization in plant genome editing.

Keywords: CRISPR/Cas; base editors; multiplex CRISPR; plant genome editing; prime editors.

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Conflict of interest statement

Declaration of interests Y.Q. is a consultant for Inari Agriculture and CTC Genomics. The remaining authors declare no conflict of interests.

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Construct design for CRISPR/Cas-based genome editing in plants

Affiliations

Construct design for CRISPR/Cas-based genome editing in plants

Authors

Affiliations

Abstract

Conflict of interest statement

Publication types

MeSH terms

LinkOut - more resources

Full Text Sources

Other Literature Sources