Abstract:

Recombinant DNA technology was just an idea a century ago that desirable characteristics in living bodies could be improved by controlling the expression of target genes. However, in recent years, this field has made significant contributions to human progress. This technology allows for the safe, affordable, and sufficient production of critical proteins for health problems and dietary purposes. This technology has multidisciplinary applications and the potential to improve important aspects of life, such as health, food resources, and resistance to diverse adverse environmental effects. Genetically modified plants, particularly in agriculture, have increased resistance to harmful agents, increased product yield, and demonstrated increased adaptability for better survival.

Introduction:

By developing new vaccines and pharmaceuticals, recombinant DNA technology is helping to improve health conditions. Treatment strategies are also being improved through the development of diagnostic kits, monitoring devices, and novel therapeutic approaches. One of the most prominent examples of genetic engineering in health is the synthesis of synthetic human insulin and erythropoietin by genetically modified bacteria [3] and the production of new types of experimental mutant mice for research purposes. Similarly, genetic engineering approaches have been used to address environmental issues such as converting wastes into biofuels and bioethanol [4-7], cleaning up oil spills, carbon, and other toxic wastes, and detecting arsenic and other contaminants in drinking water. Genetically modified microbes can also be used for biomining and bioremediation.

The invention of recombinant DNA technology revolutionised biological research and resulted in a series of dramatic changes. The pharmaceutical products synthesised using recombinant DNA technology completely changed the human life in such a way that the United States Food and Drug Administration (FDA) approved more recombinant drugs in 1997 than in the previous several years combined, including anaemia, AIDS, cancers (Kaposi’s sarcoma, leukaemia, and colorectal, kidney, and ovarian cancers), hereditary disorders (cystic fibrosis, familial hypercholeste.

Various factors endanger human life, such as food scarcity, which leads to malnutrition, various types of lethal diseases, environmental issues caused by rapid industrialization and urbanisation, and many others. Traditional strategies have been replaced by genetic engineering, which has the greater potential to overcome such challenges. The current review summarises the major challenges that humans have faced and discusses the role of recombinant DNA technology in overcoming these issues. In line with this, we have detailed the limitations of genetic engineering as well as possible future directions for researchers to overcome such limitations by modifying current genetic engineering strategies.

Recombinant DNA Technology:

Recombinant DNA technology entails modifying genetic material outside of an organism in order to obtain enhanced and desired characteristics in living organisms or their products. This technology entails inserting DNA fragments from various sources with a desired gene sequence into an appropriate vector. Manipulation of an organism’s genome occurs through the introduction of one or more new genes and regulatory elements, or by decreasing or blocking the expression of endogenous genes via recombining genes and elements. Enzymatic cleavage is used to obtain different DNA fragments for specific target sequence DNA sites, followed by DNA ligase activity to join the fragments to fix the desired gene in the vector. The vector is then introduced into a host organism, which is grown in culture to produce multiple copies of the incorporated DNA fragment, and clones containing a relevant DNA fragment are chosen and harvested.

Current Research:

Recombinant DNA technology is a rapidly expanding field, with researchers all over the world developing new approaches, devices, and engineered products for use in a variety of industries, including agriculture, health, and the environment. In comparison to regular human insulin, Lispro (Humalog) is a highly effective and rapidly acting recombinant insulin [3]. Similarly, Epoetin alfa is a novel and well-known recombinant protein that can be used to effectively treat anaemia.

Clustered regularly interspaced short palindromic repeats (CRISPR), a recent advancement in recombinant DNA technology, has provided solutions to a variety of problems in various species. This system can be used to specifically destroy genes in human cells. Gene activation, suppression, addition, and deletion in human cells, mice, rats, zebrafish, bacteria, fruit flies, yeast, nematodes, and crops demonstrated the technique’s viability. With CRISPR, mouse models can be managed for studying human diseases, where individual gene studies become much faster and gene interactions become simple by changing multiple genes in cells.

CRISPR-Cas, the only adaptive immune system in prokaryotes, contains a CRISPR genomic locus with short repetitive elements and spacers (unique sequences). The AT-rich leader sequence precedes the CRISPR array, which is flanked by cas genes that encode Cas proteins [32, 33]. Catalases cas1 and cas2 in E. coli promote the formation of new spacers via complex formation. Because the target sequence is not chosen at random, a photo-spacer adjacent motif (PAM) is required for interference and acquisition. After CRISPR array transcription into long precursor crRNA, memorization of the invader’s sequence begins. Target is degraded during the final stages of the immunity process due to interference with invaded nucleic acids. The system is prevented from self-targeting by specific recognition.

Future Prospects:

Recombinant DNA technology as a gene therapy tool is a source of prevention and cure for acquired genetic disorders. The development of DNA vaccines is a novel approach to providing immunity against a variety of diseases. The DNA delivered in this process contains genes that code for pathogenic proteins. In clinical trials, human gene therapy is primarily used to treat cancer. The majority of research has concentrated on high transfection efficacy in gene delivery system design. Transfection for cancer gene therapy with low toxicity, such as in the cases of brain cancer, breast cancer, lung cancer, and prostate cancer, is still being researched. Gene therapy is also being considered for renal transplantation, Gaucher disease, haemophilia, Alport syndrome, renal fibrosis, and other diseases.

Conclusion:

Recombinant DNA technology is a significant scientific advancement that has made human life much easier. It has advanced strategies for biomedical applications such as cancer treatment, genetic diseases, diabetes, and several plant disorders, particularly viral and fungal resistance, in recent years. The role of recombinant DNA technology in cleaning up the environment (phytoremediation and microbial remediation) and improving plant resistance to various adverse acting factors (drought, pests, and salt) has been widely recognised. It made significant improvements not only in humans but also in plants and microorganisms. The challenges in improving products at the gene level sometimes face serious difficulties that must be addressed for the future of recombinant DNA technology.