MicroRNA Transformed Genetically Modified Crops and their Effect on Human Health.


 GM crops or Genetically Modified crops are attracted a wide range of media attention in recent years and continues to do so. Media given awareness about the genetically modified crops to public. They reported the uses and drawbacks of the GM crops. The technique offers with regards to the range of advantages of the use of genetically modified crops. In the Pioneer stage of the production of GM crops, two different sectors of concern have been evolved, which includes impact on the agriculture and dangerous to human life. Safety of the eatables have a vital role in the world. The issue can be reduced by enhancing the productivity and quality of the crop. Genetic manipulation technology depends on the MiRNA, it is one of the main problem-solving methods, which influence the environmental product formation through improving major rules used for miRNAs modification and its objectives in GM plants, it contains constitutive, induction to stress, or specific tissue expression of micro RNAs or their aim, RNA gene silencing mechanism, micro-RNA-resistant target and gene expressions. Genetically Modified Organisms is one of the major focuses in biomedical research from 1980s. Since, Genetically Modified models with animal enable researchers for treatment of human genetic diseases. Genetically Modified microorganisms, crops, and animals are used for the production of drugs that are complex by which helps the generation to vaccines that are cheaper. However, this article+ more focused on the human health associated with the genetically modified foods and role of miRNAs in respected to GM food products.


Introduction
200,000 natural substances are produced and accumulated by plant crops [1]. That can be used as eatables, avours, medicines, colouring dyes etc. Some of the compounds such as alkaloids, terpenoids, steroids, polyketides, phenylpropanoids and avonoids were obtained from plant variety which is economically important. GM crop methods involves the transfer of genes of interest from one plant to another or from one plant kingdom to another target plant. Crops are very important part of the environment. Which utilised by humas for shelter, pharmaceuticals and also for eatables. The major impact on our ecosystem caused by humas. Human exercises incorporate abiotic stresses like dry season, soil harmfulness, environmental alteration, and biotic dangers like creepy crawlies, herbivores, microbial microorganisms, and so on the expansion in the populace on the planet with the associative expansion popular authorization of plant items an expansion in harvest capitulate. One such strategy includes decreasing the yield punishments by planning and receiving earth cordial harvest insurance measures [2][3][4][5]. These riboregulatory can hence ll in as particles that can be controlled to support promoting plant e ciency. This audit gives a short portrayal of micro-RNA formation and revelation. It additionally examines the job of microRNAs as potential quality controllers in grains, vegetables, tubers, natural products, sources of biofuel, refreshments, and crops with bre dependent on the latest distributions. The procedures depend on miRNA utilized for enhancing nancially signi cant plants are additionally depicted.
In recent studies shows that microRNAs plays an important role in life of plant, especially environmental stress response, development and architecture of plant using microarray and high-throughput sequencing methods [6-9].

Genetically Modi ed Plants : An Introduction
Verity of methods exists for the manufacturing of Genetically Modi ed crops. Two methods are commonly suggested for the production of GM crops. The 'gene gun', which transfer the DNA coated on a microparticle in to a cell of a crop. DNA should be transferred into target plant cell that will propagate to produce GM plants by tissue culture mechanism [10]. The main concepts of the protocol are as follows The transformed cells are nding the help of selectable markers.
Delivery of desired DNA into the crops genetic material The chances of high mutations in Genetically Modi ed crops compared to non-Genetically Modi ed crops are used in rearrangement and synthesis of DNA along with addition of new genes.
To enhance the delivery of desired DNA into plant cell genome, a selectable marker gene was provided.
e.g., Antibiotic resistance, is transfer of desired DNA to allow discrimination of genetically modi ed tissue and development of genetically modi ed crops. There will be a problem due to consumption of genetically modi ed plant. When it consumed by human that will affect the micro ora in the human gut and the microorganisms that are resistant to antibiotic in the soil. There by the antibiotic resistance gene in the bacteria will spread in the bacterial colonies. As a result of production of GM crops, that will have large mutation than the original plant [11]. Tissue culture techniques that can cause mutations in plants, causes soma clonal variation and rearrangements of DNA may occur in transgene, where the plants can produce toxins in theoretical aspects [11] 2.1 Genetically Modi ed plants act as a resource of foods Collection and generation of eatable yields, bers and another product is one of the methods used in past days. The plants that is genetically manipulated plants can be used as a food, and which is also helps in the production of oils (extractable) or can give proteins system expression. There are multiple problems to solve when transfer from studies to wet lab work [12]. Direct application, shows the effects and market for the particular character of the target and it will helps for a better knowledge of metabolism for gene regulation to new product that is applied. The information regarding expression system in particular tissue or organ or organelle, in various stages of progression can be known, for getting largest level of accumulation. For con rming the safety measures of new product consumption there must be a constant or predictable accumulation level is required for nutraceuticals [13,14]. Golden Rice, is the second generation of genetically modi ed foods, gives a better research study for individual concerns. [15].

Future of Genetically Modi ed plants
Acceptance of genetically modi ed plants by public is the main hurdles, commercially. Because of the safety related to primary generation of modi ed plants, it will take much time for getting public acceptance [16]. Metabolomics is one of the feasible solutions for analysis of comprehensive metabolite [17,18]. Well known cells like; transgenic animals, yeast, bacteria, and mammalian cell culture were included in the recombinant protein expression systems. The main factors implicated for commercial applications are, the expenses during operation and culture [19]. While generating recombinant proteins such as antibodies and vaccines, folding of proteins and manipulation of post-translational product that is glycosylated, are the main problems. It shows that plants are better systems for producing protein products that are effective and also active [19] 3. . Those challenging traits can be unraveled by the regulated networks of miRNA or advances in miRNAs functional analysis [7]. miRNAs can represent themselves as a gene reservoir, which can act as a new candidate for manipulation of challenging traits [7,8,24]. The elucidation of miRNA directed interactions will leads to the detection of much useful transcription factors and well known of the mechanisms of several processes in which transcription factors are implicated.

MiRNAs and its activity for improving crop yield
The research shows that microRNAs act a crucial effect in responses to different stresses including infections of bacteria and fungus, de ciency of nutrients, salt, drought, cold and temperature. The possibility of the important activity recognized by the detection of multiple miRNAs that response to stress produced by various tools of bioinformatics; analysis and database sequence prediction, analysis of microarray [6-9, 22, 26, 27]. So, confusions about molecular level mechanism, in uence of microRNA in accordance to plant stress, detection of regulation network of gene via miRNA and characterization of particular miRNA were cleared utilizing the research information. For developing new varieties of stress resistant having good activity during various environmental stresses, some transgenic methods are used in crop species for over expressing or knock out/down genes of particular miRNAs. So, these modi cation of micro-RNA in transgenic plants leads to make better results in biomass and seed yield, it agricultural productivity of agriculture. The review is separated into 5 parts, which includes regulation networks of gene via miRNA, and progress of the study about the modi cation of miRNAs expression in transgenic plants for improving agricultural production.

miRNA Target Screening and Prediction Methods
Identi cation of target genes of miRNAs of plants, utilizing tools of bioinformatics by experimental methods [44][45][46]. With the help of complementarity scoring and analysis of secondary structure are: psRNATarget; TAPIR, miRTour and miRTarBase, the experimentally validated aimed relation of miRNA database were utilized for target identi cation of miRNA [48, 46, 47].
Real-time PCR is used for evaluating target mRNAs expression level, the cutting site of target can be structured with the help of 5′-Rapid Ampli cation of cDNA Ends or RACE. Degradome sequencing method was progressed, recently. So, this one is the manipulated version of 5′-Rapid Ampli cation of cDNA Ends utilizing a high-throughput, deep sequencing method [49,50,45].

Functional Roles of miRNAs
Arabidopsis plant and others are shows that micro RNAs are implicated in multiple procedures like growth and development, genome integrity maintenance, transduction of signal, signalling pathways of hormone, homeostasis of hormone, innate immunity, and abiotic and biotic stress response [36,7,51,47]. Recent study reported that micro RNAs can be used to reschedule metabolism of cells at the time of plant attacked by pathogen. It leads to changes in dynamic to the micro transcriptome connected with regulation of differential transcription with the help of immunity and basal resistance [53]. Proteins or enzymes were coded by gene transcript in metabolic process.

Rice (Oryza sativa)
In world, half of the populations are using rice as their rst resource of food. There are some factors that affect growth and yield of the crops, those are drought, salinity, ice cold, heat and de ciency in nutrients. From the library of cDNA of rice, 20 miRNAs were primarily recognized in 2014 and their forecasted genes of target were concerned in transfer, resistance to disease, transcription, metabolism etc. [54]. In another research, 35 rice miRNAs were recognized in the upcoming year, from that 14 of them are new and target genes were included in various physiological procedures in rice were also forecasted [55]. 592 miRNAs sequences contained in version 21 were recognized in miRNAs of multiple rice. High tillering and early blossoming are the two phenotypes which are important for productivity and it was showed by OsmiR393, plants that is overexpressing [56]. For responding to heat stress and adaptation of rice by Lascorbate oxidase (OsLAC) expression, which is controlled by OsmiR397, reported as a responsive miRNA for high-temperature [57,58] conservation of 20 families such as miRNA156/157, miR159, miR160, miR164, miR165/166, miR167, miR168, miR169, miR170/171, miR172, miR319, miR390, miR393, miR396, miR397, miR399, and miR408, and in which 23 of them are noval [59]. The following proteins like squamosa enhancer binding proteins, scarecrow-like blue copper proteins are included for the growth of wheat crop as well as its various physiological procedures, so these are directed by the above miRNAs [59]. In miRbase version 21 containing 116 miRNAs sequences have identi ed more miRNAs of wheat. Characterization of tissues of wheat and 323 new miRNAs included in 276 families were conducted a wide survey with their related target genes very recently. The above miRNAs plays a signi cant part in progress of grain and also it will expressed in grain[60].

Maize (Zea mays)
In world, maize is secondary crucial crop, that is utilized for feed as well as forage resource for fuel production in ethanol. Maize will use as a model plant in studies due its agricultural and economic importance

vine of Grape (Vitis vinifera L.)
Grapevine having best nutritional and processing qualities, so it can be cultivated in worldwide and it is also economically important fruit crop. Genome sequencing of grapevine was completed, so scientists were much interested to miRNAs investigation [68,69]. To miRbase 21, 349 miRNAs of grapevine are settled. For understanding the functions of already detected miRNAs, there is a prediction of known miRNAs 112 aimed genes and novel grapevine-speci c miRNAs having 44 target genes. In one of the researches, 25 grapevine miRNA families have to aim total 134 target genes. So, these genes will encode transcription factors that are included in growth of plant, development of plant, and its phase alters from vegetative to reproductive. ATP sulfurylase/APS kinase and ATP synthase were encoded by target genes which plays an important role in various procedures of metabolism. Those target genes involved in resistance to disease, response to immunity, response to stress, and signaling transduction [70]. Grape vine implicated in biotic and abiotic stresses responses; a study suggested that cold-inducible grapevine miRNAs play a major role in response to cold stress [71].

Apple (Malus domestica Borkh)
It is another economically important tree of fruit, and consumed widely. It contains avonoids and compounds of phenol, that plays an important function in decreasing chronic disease risk in humans [72]. From apple, identi cation and characterization of 413 miRNAs in miRBase version 21. Identi cation of 146 microRNAs from M. domestica (cv. Golden Delicious) utilizing bioinformatic analysis and Ribonucleic Acid library sequencing methods. The research suggested that new miRNA of apple, Md-miRLn11, controls the NBS-LRR protein expression at the time of infection of pathogen which leading to plant bacterium resistance [73]. Some of the miRNAs are implicated in re blight resistance, caused by Erwinia amylovora, bacterial disease.

Tomato (Solanum lycopersicum L.)
Tomato is widely consuming and commonly using vegetable and also it can utilize as a experimental plant model for the research of ripening and senescence which attracted by scientists for detecting function of miRNA at the time of development of fruit by aiming genes implicated in pathway of ethylene and ripening of fruit [74]. Small Ribonucleic Acids from fruit and leaf of tomato were detected, majority of them plays a crucial role in development of fruit. miR1917, a key negative regulator of ethylene responses, acts on ripening of tomato [75]. In a research, studied about the defense mechanism of microRNAs in tomato against Fusarium oxysporum. It demonstrates that miR482/2118 inhibits genes of nucleotide-binding site, which cause resistance to F. oxysporum [76]. Before, it was suggested that 6. Genetically Modi ed Foods: Bene ts 6.1 Resistance to pest The plants or crops that are modi ed using genetic engineering techniques has modi ed DNA that show resistance to pest and agents that are speci c and harmful to plants which ensures the growth improvement of crops and helps farmers for good yield. Mutagenesis and GE varies as the exposure to chemicals and radiation contribute mutagenesis that cause stable changes that are not speci c [77] Other methods in breeding such as Selective plant or animal breeding or soma clonal variation are also used by human to for modi cation of food, mainly by addition of a new trait that is not present naturally which shows characteristics of pest resistance, worse diseases and also certain nutrient or drug agent that are bene cial. Genetically modi ed foods such as Bt. Corn doesn't need the addition of pesticides as it modi ed in such a way which is cost effective during the time to harvest [78]

Tolerance to Herbicide and Resistance to Disease
The environmental hazards caused by herbicides can be controlled by plants or crops that are engineered to resist herbicide. [79][80][81] Genetically-engineered crops designed for herbicide resistance provide a positive environmental aspect with the limited use of herbicides [79][80][81], in turn reducing its production expense and limiting the leaching of agricultural waste. A number of plant diseases are triggered by the infection of various viruses, fungi and bacteria. Plant scientists points out the fact that genetically modi cation likely tends to have least unintended changes than the conventional bred crops, which may be inferred from the results of a detailed crop composition pro le. In advancement, genetically engineered plants which resist the speci c plant infections are being designed by plant biotechnologists [82].

Cold, Drought and Salinity Tolerance
A recent report shows that plants such as tobacco and potato has been successfully introduced with an antifreeze gene from a cold-water sh, resulting them to withstand the cold temperatures, and generally killing unmodi ed seedlings [82].The rise in global population has resulted the increase in demands for land in housing rather than food cultivation, leading farmers to cultivate in previously unsuitable localities for crops. The modi ed plants aimed to survive periods of drought, cold or high salinity in soil and groundwater would enable to cultivate on such uncongenial dwellings [83].

Nutrition Properties
Undernourishment is commonly prevalent in the third world countries, as the poor rely on a single crop such as rice, as their major staple diet. Unfortunately, rice does contain all the essential nutrients in adequate amounts to tackle malnutrition. So, if rice could be modi ed to incorporate all the additional vitamins and minerals with adequate nutraceutical signi cance, nutrient limitations could be satis ed. A study conducted at the Swiss Federal Institute of Technology for Plant Sciences lead to the development miR159/319 and miR172 might contributes its role in pathogenesis of viruses in tomato against the curl virus of Tomato leaf, chilling and freezing like Cold stresses.
of 'golden rice', incorporating a high content of beta-carotene(Vitamin A) [84,85].Further improvements on the golden rice are also studied, having increased iron content.

Pharmaceutical Characteristics
The production of medicines and vaccines are often costly, and certain times require speci c storage condition. Working on this speci c line of action led to the way in developing certain edible vaccines in potatoes and tomatoes [86],probably compromising with easier way to ship, store and administer linkage maps for cows, pigs and sheep explaining chromosomal zones for nancially signi cant characteristics will observably pay to improved worth and amounts of meat. Quality innovation is wealthy in livestock creation and in progress of value and amount attributes [91]. Genetic technology persuades the creations, more nutrient supplement and animals' prosperity. These singularities can be updated straightforwardly by quality exchange or utilizing development hormones, immunizations, antibodies, resistance energizers and antiallergy DNA made by technology associated with genetics. Quality exchange is foreseen to propel those creation characteristics in animals, which are ailing inherited, for instance of forestalled piglets [92] depicted transgenic crops, that delivered immunizations in which animals overwhelmed by rummage, are framed. The quality of encounter enables rearing of animals impenetrable to infections. Immunization for invulnerable maiming of animals, easy in male animals and debilitates decisiveness while female animals are liberated from negative impacts of oestrus, unquestionably in uences the monetarily critical quality remains adaptation. All things considered; transgenic milk can be utilized as: (a) Food for wide use; (b) crude materials for milk items; (c) nourishment for babies; (d) wellspring of organically dynamic substances for drug industry. Indeed, even non-protein mixes of human milk, similar to oligosaccharides, are incredibly esteemed in milk of transgenic animals. Caseins and lactoglobulins are created distinctly during lactation period. Qualities from referenced mixes are utilized for transgenic milk creation, being used for cheese creation and for assistant to human milk with the end goal of baby sustenance depicted on wide utilization of ox-like development hormone (somatotropin) in steers to raising creation of both, meat and milk [93]. Today there is still a lot to learn about science just as hereditary designing consider with human milk, and thus there is a lot to chip away at the best arrangement of baby formulae taking into account overhauling the human milk's quality utilizing an ever-increasing number of exact works on with respect to hereditary change.

Conclusion And Future Perspectives
The research concluded that, currently miRNAs are taken as the most crucial regulators of gene. Until now, there is a signi cant progress has been made plant miRNAs analysis and characterization, with large amount of research conclusions in miRNA's important function in crops. The review summarizes that several ndings that contribute miRNA and its agricultural and economic importance [7]. The small nucleic acids regulate different environmental stresses also in uences the growth of plants, the signaling of hormones, reproductive and vegetative phase changes along with the responses in homeostasis and signal transduction pathways. A few studies concluded that miRNA of crops shows in molecular regulations of defense and immune responses of plant. For the enhancement in the agricultural properties of plants and crops, the genetic engineering targets the modi cation of miRNA [35,27].
The manipulation of miRNA and its expression levels focus on the responses of plants against pathogens, parasites and environmental stress which enhances it growth, for this purpose several transgenic approaches such as over expression, expression speci c to tissues, stress-induced, arti cially target mimic etc., are focused on important miRNAs [47].
In such cases the miRNA act as positive regulators of stress that suppress the corresponding messenger RNA which leads to the improvement of plants which causes an undesirable effect in the native target genes. In cases when the target genes show a desirable effect then the miRNA acts as negative stress regulators which leads to over expression or arti cial target mimics of the target genes [94]. Even though its feels like successful the practical application of the miRNA crops is challenging in agronomics as the complexity of the multiple genetic traits requires tuning of different genes in various stages of the development of plants. For the future crop improvement, the above-mentioned methodologies and a detailed analysis on the miRNA-based regulation of target in crops and plants will bene t the future strategies and design. The over expression of the miRNA and the knockout of important genes leads to same phenotypes and can lead to the reduction of the miRNA is another major [95].
The miRNA activity regulates and act as a e cient tool to relate the activities if microRNA families and engineering of plants gene expression and this experiment offer a chance that can alter the sequence of decoy sites of miRNA that enhances the inactivation of miRNA and its targets also affects the production of plant phenotypes. But the major challenges faced is of false positive outcome by off-type effects of due to inactivation of miRNA, the action and communication between the miRNA and its decoy are very complex and that can also lead to plants losing its stability [96].
The latest invasion of biochemistry and physiology has a special focus in GE and transgenic technology has several uses in production of microorganism which includes plants, microorganism and animals. The augmentation via genetic alteration has in uenced several crops. The foods that are genetically modi ed have much bene ts such as tolerance towards salinity, resistance towards insects and high harvest. Genetically modi ed foods have numerous acts on health on human, these foods have both good and bad effects, which can be direct effect on the human that is consuming it or can be wider impacts such as in food chain that includes several other organisms.
For instance, if the expression of a miRNA or the targeted gene is changed, it may result in unacceptable pleiotropic changes in the developmental and morphological characteristics of that particular plant species. Hence, it is important to realize the miRNA regulation method in growth and development of plant or its reaction to different stresses. This will ease the pattern of procedures which are satisfying and thus results in the wish for traits but with limited trade-offs when it comes to modi es crops.