International Research Journal of Engineering and Technology (IRJET)
e-ISSN: 2395-0056
Volume: 11 Issue: 03 | Mar 2024
p-ISSN: 2395-0072
Mechanisms and Applications of CRISPR mediated Gene Silencing Ranjanee Chattopadhyay*1, Rajpal Srivastav1 1 Amity University Uttar Pradesh, Noida, Uttar Pradesh, India
---------------------------------------------------------------------***--------------------------------------------------------------------is a new and intriguing technique which has fascinated the Abstract – Genetic engineering has fascinated the world world with its immensity of opportunities.
with its various techniques to manipulate the genes of organisms. The gene silencing technique, known as CRISPR, has now come to the limelight as it has successfully mitigated the long-standing limitations of the previous gene editing techniques. It is cost-effective, faster, accurate, and highly precise. It can successfully suppress the pathogenic genes, thus reforming the agricultural and healthcare sector. The advanced CRISPR-Cas9 mediated gene editing has become a leading tool in the therapeutics of genetic diseases and disorders. In this article, the mechanism of the CRISPR-Cas9 gene silencing technique and its recent trends in crop improvement and therapeutics have been highlighted along with regulatory, ethical concerns and prospects.
1.1. HISTORY
CRISPR, which stands for, Clustered Regularly Interspaced Short Palindromic Repeats, was first discovered by Ishino in the year 1987 in Osaka University, Japan, while working with DNA sequences form Escherichia coli bacteria. The exact function and significance were unknown at that time [5] (Ishino Y et al, March 2018). Significant information regarding CRISPR-Cas9 was obtained in 2007 by two French scientists, Rodolphe Barrangou and Philippe Horvath, who had been working with yoghurt cultures of bacteria Streptococcus thermophilus for the Danish company Danisco [6] (Gostimskaya I, August 2022).
Key Words: CRISPR, Cas9 protein, Gene editing, Genetically Modified Crops (GMCs), Gene knockdown, Crop improvement, Therapeutics, Epigenetic modification
The first demonstration of the mechanism of CRISPR-Cas9 was made in 2008 by Eugene Koonin and colleagues, National Center for Biotechnology Information, Maryland [7] (Ratan ZA et al, Feb 2018). Slowly after that, many more publications were made, and more understanding of this mechanism has been made. In recognition of the importance of CRISPR-Cas9,the Nobel Prize in chemistry was awarded to Jennifer Doudna and Emmanuel Charpentier in 2020 [8] (Uyhazi KE et al, Jan 2021).
1.INTRODUCTION Gene silencing, also known as epigenetic modification of gene expression, is a molecular biological process which involves gene silencing and/or gene knockdown and aims to control diseases by eliminating or suppressing the targeted gene from the genome. Since genes can be regulated either at the transcriptional level or at post-transcriptional level, gene silencing can be induced in both levels.
CRISPR-Cas9 is a revolutionary genome-editing and gene silencing technology. It has high precision, is highly flexible and allows for precise modification of DNA sequences and modification & regulation of genes. It consists of a negative feedback mechanism which knocks down gene expressions and takes part in defence mechanisms. It is better than the previously known genetic engineering techniques due to the fact that it allows removal & introduction of more than one target gene at a time, is much less time-consuming and less costly [9] (Uddin F et al, Aug 2020). Also, it is not speciesspecific and hence, a wide range of organisms which were previously resistant to these genetic engineering techniques can now be explored.
Researchers are studying gene silencing due to its potential to develop genetically modified crops(GMCs) and plants with better yield and higher resistance to target pests and also protect nature [1] (Zhang Y et al, May 2017). In the future, it may allow opportunities such as human gene therapy [2] (Piergentili R et al, April 2021), programmable RNA targeting, more advancements in agriculture and the health sector, a probable permanent cure for diseases which are incurable to date. It may also produce therapeutics to battle cancer, infectious diseases and neurodegenerative disorders [3] (Pickar-Oliver A and Gersbach CA., Aug 2019) and more [4] (Grand Moursel L et al, April 2021). Gene silencing is a highly specific and targeted procedure. With the help of this revolutionary technique, a variety of characteristics of both plants and animals can be altered to a great extent, which will itself eradicate a lot of problems related to pest control, crop yield, food scarcity, human health and well-being. CRISPR-Cas9 mediated gene silencing
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