Compendium of Plant-Specific CRISPR Vectors and Their Technical Advantages
CRISPR/Cas mediated genome editing is a revolutionary approach for manipulating the plant genome. However, the success of this technology is highly dependent on selection of a specific vector and the other components. A plant-specific CRISPR/Cas vector usually consists of a <i>Cas</i> ge...
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MDPI AG
2021-09-01
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| Series: | Life |
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| author | Anshu Alok Hanny Chauhan Santosh Kumar Upadhyay Ashutosh Pandey Jitendra Kumar Kashmir Singh |
| author_facet | Anshu Alok Hanny Chauhan Santosh Kumar Upadhyay Ashutosh Pandey Jitendra Kumar Kashmir Singh |
| author_sort | Anshu Alok |
| collection | DOAJ |
| description | CRISPR/Cas mediated genome editing is a revolutionary approach for manipulating the plant genome. However, the success of this technology is highly dependent on selection of a specific vector and the other components. A plant-specific CRISPR/Cas vector usually consists of a <i>Cas</i> gene, target-specific gRNA, leader sequence, selectable marker gene, precise promoters, and other accessories. It has always been challenging to select the specific vector for each study due to a lack of comprehensive information on CRISPR vectors in one place. Herein, we have discussed every technical aspect of various important elements that will be highly useful in vector selection and efficient editing of the desired plant genome. Various factors such as the promoter regulating the expression of Cas and gRNA, gRNA size, Cas variants, multicistronic gRNA, and vector backbone, etc. influence transformation and editing frequency. For example, the use of polycistronic tRNA-gRNA, and Csy4-gRNA has been documented to enhance the editing efficiency. Similarly, the selection of an efficient selectable marker is also a very important factor. Information on the availability of numerous variants of Cas endonucleases, such as Cas9, Cas12a, Cas12b, Casɸ, and CasMINI, etc., with diverse recognition specificities further broadens the scope of editing. The development of chimeric proteins such as Cas fused to cytosine or adenosine deaminase domain and modified reverse transcriptase using protein engineering enabled base and prime editing, respectively. In addition, the newly discovered Casɸ and CasMINI would increase the scope of genetic engineering in plants by being smaller Cas variants. All advancements would contribute to the development of various tools required for gene editing, targeted gene insertion, transcriptional activation/suppression, multiplexing, prime editing, base editing, and gene tagging. This review will serve as an encyclopedia for plant-specific CRISPR vectors and will be useful for researchers. |
| first_indexed | 2024-03-10T06:26:42Z |
| format | Article |
| id | doaj.art-2b25064012d745fe80146a264ce96429 |
| institution | Directory Open Access Journal |
| issn | 2075-1729 |
| language | English |
| last_indexed | 2024-03-10T06:26:42Z |
| publishDate | 2021-09-01 |
| publisher | MDPI AG |
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| series | Life |
| spelling | doaj.art-2b25064012d745fe80146a264ce964292023-11-22T18:52:16ZengMDPI AGLife2075-17292021-09-011110102110.3390/life11101021Compendium of Plant-Specific CRISPR Vectors and Their Technical AdvantagesAnshu Alok0Hanny Chauhan1Santosh Kumar Upadhyay2Ashutosh Pandey3Jitendra Kumar4Kashmir Singh5Department of Biotechnology, Panjab University, Chandigarh 160014, IndiaDepartment of Biotechnology, Panjab University, Chandigarh 160014, IndiaDepartment of Botany, Panjab University, Chandigarh 160014, IndiaNational Institute of Plant Genome Research, New Delhi 110067, IndiaDepartment of Agronomy and Plant Genetics, University of Minnesota, Saint Paul, MN 55108, USADepartment of Biotechnology, Panjab University, Chandigarh 160014, IndiaCRISPR/Cas mediated genome editing is a revolutionary approach for manipulating the plant genome. However, the success of this technology is highly dependent on selection of a specific vector and the other components. A plant-specific CRISPR/Cas vector usually consists of a <i>Cas</i> gene, target-specific gRNA, leader sequence, selectable marker gene, precise promoters, and other accessories. It has always been challenging to select the specific vector for each study due to a lack of comprehensive information on CRISPR vectors in one place. Herein, we have discussed every technical aspect of various important elements that will be highly useful in vector selection and efficient editing of the desired plant genome. Various factors such as the promoter regulating the expression of Cas and gRNA, gRNA size, Cas variants, multicistronic gRNA, and vector backbone, etc. influence transformation and editing frequency. For example, the use of polycistronic tRNA-gRNA, and Csy4-gRNA has been documented to enhance the editing efficiency. Similarly, the selection of an efficient selectable marker is also a very important factor. Information on the availability of numerous variants of Cas endonucleases, such as Cas9, Cas12a, Cas12b, Casɸ, and CasMINI, etc., with diverse recognition specificities further broadens the scope of editing. The development of chimeric proteins such as Cas fused to cytosine or adenosine deaminase domain and modified reverse transcriptase using protein engineering enabled base and prime editing, respectively. In addition, the newly discovered Casɸ and CasMINI would increase the scope of genetic engineering in plants by being smaller Cas variants. All advancements would contribute to the development of various tools required for gene editing, targeted gene insertion, transcriptional activation/suppression, multiplexing, prime editing, base editing, and gene tagging. This review will serve as an encyclopedia for plant-specific CRISPR vectors and will be useful for researchers.https://www.mdpi.com/2075-1729/11/10/1021CRISPR/Cas9Csy4-gRNAprime editingmultiplexingbase editing |
| spellingShingle | Anshu Alok Hanny Chauhan Santosh Kumar Upadhyay Ashutosh Pandey Jitendra Kumar Kashmir Singh Compendium of Plant-Specific CRISPR Vectors and Their Technical Advantages Life CRISPR/Cas9 Csy4-gRNA prime editing multiplexing base editing |
| title | Compendium of Plant-Specific CRISPR Vectors and Their Technical Advantages |
| title_full | Compendium of Plant-Specific CRISPR Vectors and Their Technical Advantages |
| title_fullStr | Compendium of Plant-Specific CRISPR Vectors and Their Technical Advantages |
| title_full_unstemmed | Compendium of Plant-Specific CRISPR Vectors and Their Technical Advantages |
| title_short | Compendium of Plant-Specific CRISPR Vectors and Their Technical Advantages |
| title_sort | compendium of plant specific crispr vectors and their technical advantages |
| topic | CRISPR/Cas9 Csy4-gRNA prime editing multiplexing base editing |
| url | https://www.mdpi.com/2075-1729/11/10/1021 |
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