Recent Advances in Plant Biotechnology and Genetic Engineering Applications in Agriculture (original) (raw)
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CRISPR/Cas9 System, an Efficient Approach to Genome Editing of Plants for Crop Improvement
RNA-Based Technologies for Functional Genomics in Plants
Crop production in agriculture is affected by plant genetic background and environmental factors. The application of modern molecular biology tools to conventional plant breeding approaches has facilitated the plant genetic improvement attempts. After the extensive employment of recombinant DNA technology in diverse plant species and despite achievements, the use of transgenic crops has been encountered with public concerns due to the presence of transgenes. The advent of sequence-specific nuclease-based editing technologies especially clustered regularly interspaced short palindromic repeat-associated protein system (CRISPR/Cas) has opened a promising avenue in genetic engineering of plants. The importance of this approach is emphasized since it is a simple and robust tool, moreover, nontransgenic mutants can be selected in later generations. Following the successful use of the CRISPR/Cas9 editing tool in model plants, the applications of this system have been increasingly reported in different plant species. This chapter reviews the contribution of the CRISPR/Cas9 system in the development of genetically modified crops with improved yield, nutritional value, and response to biotic and abiotic stress factors.
The CRISPR/Cas Genome-Editing Tool: Application in Improvement of Crops
Frontiers in Plant Science, 2016
The Clustered Regularly Interspaced Short Palindromic Repeats associated Cas9/sgRNA system is a novel targeted genome-editing technique derived from bacterial immune system. It is an inexpensive, easy, most user friendly and rapidly adopted genome editing tool transforming to revolutionary paradigm. This technique enables precise genomic modifications in many different organisms and tissues. Cas9 protein is an RNA guided endonuclease utilized for creating targeted double-stranded breaks with only a short RNA sequence to confer recognition of the target in animals and plants. Development of genetically edited (GE) crops similar to those developed by conventional or mutation breeding using this potential technique makes it a promising and extremely versatile tool for providing sustainable productive agriculture for better feeding of rapidly growing population in a changing climate. The emerging areas of research for the genome editing in plants include interrogating gene function, rewiring the regulatory signaling networks and sgRNA library for high-throughput loss-of-function screening. In this review, we have described the broad applicability of the Cas9 nuclease mediated targeted plant genome editing for development of designer crops. The regulatory uncertainty and social acceptance of plant breeding by Cas9 genome editing have also been described. With this powerful and innovative technique the designer GE non-GM plants could further advance climate resilient and sustainable agriculture in the future and maximizing yield by combating abiotic and biotic stresses.
CRISPR/Cas9 applications and future prospectus in crop genetic improvement
Journal of Multidisciplinary Sciences (e-ISSN: 2671-5449), 2020
The demand for food in 2050 is projected to rise as the global population crests about 10 billion people and meet the challenge, and agricultural production should be increased dramatically. Gene editing system has been revolutionized in genetics and crop improvement with the multipurpose technology of CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat). CRISPR/Cas9 system is based on the complementary of the guide RNA (gRNA) to a specific sequence and the Cas9 endonuclease activity that brought a new opportunity in agricultural research to develop innovative plant varieties with the addition or deletion of significant characters. The CRISPR/Cas9 technology of genome editing is variously applied to improve crop quality worldwide. CRISPR technology has shown great potential in manipulating genes in the plant, making the crops' quality and stability. CRISPR/Cas9 is a recent and reliable molecular scissor for genetic engineering, making the advanced revolution in life science. The review is based on the multidimensional application of the CRISPR/Cas9 system in plant nutritional improvement, enhancement of plant disease resistance, and drought-tolerant plant production. Cite this as: Das, B.D. & Paudel, N. (2020). CRISPR/Cas9 applications and future prospectus in crop genetic improvement. J. Multidiscip. Sci. 2(2), 56-66.
European Journal of Experimental Biology
Genome engineering with the RNA-guided CRISPR-Cas9 system in animals and plants is revolutionizing biology. First techniques of genome editing like zinc finger nucleases and synthetic nucleases called TALENs were a starting point but turned out to be expensive, difficult to handle and timeconsuming to engineer, limiting their widespread use, particularly for large scale, high-throughput studies. Moreover, these existing technologies depending on proteins as address labels and customizing new proteins for any new change to introduce in the DNA is a cumbersome process. Of the current generation of genome editing technologies, CRISPR-Cas9 is easier to use and more efficient and can be easily targeted to almost any genomic location of choice by a short RNA guide and has been successfully applied in many organisms, including model and crop plants. Together the system has the ability to detect specific sequences of letters within the genetic code and to cut DNA at a specific point. Simultaneously with other sequence-specific nucleases, CRISPR/ Cas9 has already breach the boundaries and made genetic engineering much more versatile, efficient and easy. There really doesn't seem to be a limit in applications of CRISPR system extendable from bacteria to complex eukaryotic organisms including plants changing the pace and course of agricultural, Biomedicine and Biotechnological research in the future. This review provides an overview of recent advances in genome editing technologies in plants, and discusses how these can provide insights into current plant molecular biology research and molecular breeding technology.
Ushering in CRISPR / Cas Mediated Genome Engineering for Crops
2019
Original Research Article The efficacy, diversity and modularity of CRISPR/Cas system are a driving force in the biotechnological revolution. Cas enzymes have been adopted as tools for manipulation of genomes in planta as a means to accelerate fundamental research and enable agricultural breakthroughs. Here, we review the working principles and components of CRISPR/Cas system for efficient gene editing in plants. We have also tabulated the recent work that has utilised CRISPR/Cas to improve economically important traits in plants. Although the apparent use of CRISPR/Cas mediated editing may make it appear as though researchers are toying with plant genomes, the cumulative power of this tool has made optimized and adaptable plant species towards permitting crucial advances in crop improvement.
CRISPR/Cas: A powerful tool for gene function study and crop improvement
Journal of Advanced Research, 2021
Background: It is a long-standing goal of scientists and breeders to precisely control a gene for studying its function as well as improving crop yield, quality, and tolerance to various environmental stresses. The discovery and modification of CRISPR/Cas system, a nature-occurred gene editing tool, opens an era for studying gene function and precision crop breeding. Aim of Review: In this review, we first introduce the brief history of CRISPR/Cas discovery followed the mechanism and application of CRISPR/Cas system on gene function study and crop improvement. Currently, CRISPR/Cas genome editing has been becoming a mature cutting-edge biotechnological tool for crop improvement that already used in many different traits in crops, including pathogen resistance, abiotic tolerance, plant development and morphology and even secondary metabolism and fiber development. Finally, we point out the major issues associating with CRISPR/Cas system and the future research directions. Key Scientific Concepts of Review: CRISPR/Cas9 system is a robust and powerful biotechnological tool for targeting an individual DNA and RNA sequence in the genome. It can be used to target a sequence for gene knockin, knockout and replacement as well as monitoring and regulating gene expression at the genome and epigenome levels by binding a specific sequence. Agrobacterium-mediated method is still the major and efficient method for delivering CRISPR/Cas regents into targeted plant cells. However, other delivery methods, such as virus-mediated method, have been developed and enhanced the application potentials of CRISPR/Cas9-based crop improvement. PAM requirement offers the CRISPR/Cas9-targted
Gene Editing and Crop Improvement Using CRISPR-Cas9 System
Frontiers in Plant Science
Advancements in Genome editing technologies have revolutionized the fields of functional genomics and crop improvement. CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat)-Cas9 is a multipurpose technology for genetic engineering that relies on the complementarity of the guideRNA (gRNA) to a specific sequence and the Cas9 endonuclease activity. It has broadened the agricultural research area, bringing in new opportunities to develop novel plant varieties with deletion of detrimental traits or addition of significant characters. This RNA guided genome editing technology is turning out to be a groundbreaking innovation in distinct branches of plant biology. CRISPR technology is constantly advancing including options for various genetic manipulations like generating knockouts; making precise modifications, multiplex genome engineering, and activation and repression of target genes. The review highlights the progression throughout the CRISPR legacy. We have studied the rapid evolution of CRISPR/Cas9 tools with myriad functionalities, capabilities, and specialized applications. Among varied diligences, plant nutritional improvement, enhancement of plant disease resistance and production of drought tolerant plants are reviewed. The review also includes some information on traditional delivery methods of Cas9-gRNA complexes into plant cells and incorporates the advent of CRISPR ribonucleoproteins (RNPs) that came up as a solution to various limitations that prevailed with plasmid-based CRISPR system.
Crispr Cas System in Plant Genome Editing a New Opportunity in Agriculture to Boost Crop Yield
2018
Clustered regularly interspaced short palindromic repeats CRISPR/Cas9 technology evolved from a type II bacterial immune system develop in 2013 This system employs RNA-guided nuclease, CRISPR associated (Cas9) to induce double-strand breaks. The Cas9-mediated breaks are repaired by cellular DNA repair mechanisms and mediate gene/genome modifications. The system has the ability to detect specific sequences of letters within the genetic code and to cut DNA at a specific point. Simultaneously with other sequence-specific nucleases, CRISPR/ Cas9 have already breach the boundaries and made genetic engineering much more versatile, efficient and easy also it has been reported to have increased rice grain yield up to 25-30 %, and increased tomato fruits size, branching architecture, and overall plant shape. CRISPR/ Cas also mediated virus resistance in many agricultural crops. In this article, we reviewed the history of the CRISPR/Cas9 system invention and its genome-editing mechanism. We also described the most recent innovation of the CRISPR/Cas9 technology, particularly the broad applications of modified Cas9 variants, and discuss the potential of this system for targeted genome editing and modification for crop improvement.
Molecular Plant, 2020
The central dogma (CD) of molecular biology constitutes the transfer of genetic information from DNA to RNA to protein. Major CD processes governing genetic flow include the cell cycle, DNA replication, chromosome packaging, epigenetic changes, transcription, posttranscriptional alterations, translation, and posttranslational modifications. The CD processes are tightly regulated in plants to maintain their genetic integrity throughout the life cycle as well as to pass the genetic material to the next generation. Engineering of various CD processes involved in gene regulation will accelerate crop improvement to feed the growing world population. CRISPR technology enables programmable editing of CD processes to alter DNA, RNA, or protein, which would have been impossible in the past. Here, an overview of recent advancements in CRISPR tool development and CRISPR-based CD modulations that expedite basic and applied plant research is provided. Furthermore, CRISPR applications in major thriving areas, such as gene discovery (allele mining and cryptic gene activation), introgression (de novo domestication and haploid induction), and application of desired traits beneficial to farmers or consumers (biotic/abiotic stress-resilient crops, plant cell factories, and delayed senescence), are described. Finally, the global regulatory policies, challenges, and prospects for CRISPR-mediated crop improvement are summarized.