The GPEL has built deep expertise in transgenesis and genome engineering, pioneering innovative approaches to manipulate the germline and to engineer cells with precision. At the core of our capabilities is a non-surgical, testicular electroporation-based platform eliminating the need for cumbersome embryo manipulation or other assisted reproductive technology-mediated transgenesis methods. In addition, we are working to develop improved methodologies for gene editing in farm animals. Through applying this integrated technology platform, we aim to develop transgenic animal bioreactors for high-value protein production, enhance productivity and desirable traits in farm animals, and advance functional genomics studies to decode gene function in physiologically relevant models spanning mice, rabbits, and large livestock.

Developing Animal Bioreactors:

The development of animal bioreactors has evolved from foundational transgenesis technologies toward advanced mammary gland engineering and biotherapeutic production. Early efforts focused on simplifying the generation of transgenic animals through non-surgical, germ-cell-mediated gene-transfer approaches, enabling efficient, rapid production of genetically modified animal models. These studies established practical alternatives to conventional embryo microinjection techniques and significantly expanded access to transgenic technology for livestock and biomedical applications.

Building upon these transgenesis platforms, the laboratory progressively shifted toward developing mammary gland-based animal bioreactors for therapeutic protein production. A key milestone was the isolation and characterization of the buffalo β-casein promoter for mammary epithelial cell-specific expression, which enabled targeted secretion of recombinant proteins into milk. Subsequently, the group demonstrated direct intraductal delivery of transgenes into mammary glands using an in-house-developed transfection agent, FA6-bPEI, bypassing germline integration and achieving expression of therapeutic proteins such as human interferon-γ in milk. This represented an important transition from conventional transgenic animal production to somatic mammary gland engineering.

The team further established this easily accessible transgenic technology in rabbits and generated a transgenic rabbit bioreactor that expresses therapeutic proteins in its milk. We have also generated a transgenic rabbit bioreactor that expresses bovine FSH in its milk. We are now working to further enhance this technology for adoption in goat models.

Extending this vision to avian systems in collaboration with ICAR-DPR at Hyderabad, we have, for the first time, isolated and characterized the ovalbumin promoter from the Indian Nicobari chicken breed, driving egg-white-specific expression of human therapeutics. Using an MMEJ-based site-specific integration at validated safe harbor loci, we have designed an expression construct that encodes human tissue plasminogen activator (hTPA), human Erythropoietin (hEPO), and bovine lactoferrin in egg whites.

Increasing Productivity in Farm Animals:

Increasing productivity in farm animals is essential to meet the growing global demand for animal-derived food products such as milk, meat, eggs, and fiber. With the continuous rise in human population and changing dietary preferences, livestock production systems must generate higher output using limited land, water, and feed resources. Improved productivity ensures better food security, enhanced farmer income, and sustainable utilization of agricultural resources. Higher productivity in livestock also improves economic efficiency by increasing feed conversion, growth rate, reproductive performance, and milk or meat yield per animal. This reduces the overall cost of production while maximizing profitability for farmers and the livestock industry. Productive animals require fewer resources to produce the same amount of output, thereby reducing greenhouse gas emissions, water consumption, and waste generation per unit of animal product. Enhancing productivity is therefore closely linked with climate-smart and sustainable livestock farming. Advances in biotechnology and genetic engineering support productivity improvements by enabling the development of disease-resistant, stress-tolerant, and high-performing animals. Technologies such as transgenesis and genome editing enable rapid genetic improvement and accelerated trait development in livestock populations. We are working at the forefront of gene editing and gene regulation to control the expression of genes that are necessary to increase productivity.  

We have adopted multiplex CRISPR-Cas9 technology to generate the myostatin MSTN gene knock-out in goat cells. We have also developed RNA interference-based strategies, named Transactivation of shRNAs in Tandem (TshRT) platform, in which multiple shRNAs can be expressed from compact minimal promoters. In this design, each TshRT unit is selectively activated by dCas9-VP64 to achieve multiplex silencing without the need to re-engineer endogenous genetic loci. Using this system, robust knockdown of diverse targets, including goat myostatin. Transgenic mice harboring a TshRT unit directed against myostatin exhibited pronounced skeletal muscle hypertrophy (double muscle phenotype), supporting in vivo efficacy and functional relevance. This modular strategy expands the toolbox for precision gene regulation and provides a promising framework for both disease modeling and the development of RNAi-based therapeutics.

Functional Genomics:

The laboratory has contributed to functional genomics by developing simplified and scalable transgenesis technologies to study gene function in vivo. The group recognized that rapid advances in genome sequencing created a major gap between identifying genes and understanding their biological roles. To address this, the laboratory focused on developing efficient animal models to functionally validate genes, transcription factors, signaling molecules, and microRNAs.

One of the major contributions was the development of non-surgical and hypotonic shock-mediated testicular transgenesis methods, which enabled rapid generation of transgenic and gene knockdown animals without complex embryo manipulation. These approaches allowed functional analysis of genes directly in living animal systems and significantly reduced the technical barriers associated with conventional transgenic technologies.

The laboratory further expanded into promoter discovery and tissue-specific gene regulation by isolating and characterizing various cell/tissue-specific promoters for controlled transgene expression. Recent work on post-meiotic germ cell-specific promoters provided important tools for studying stage-specific gene expression during spermatogenesis and reproductive functional genomics.

Using these platforms, the laboratory generated several functional genomics models to investigate reproductive biology, fertility regulation, and disease mechanisms. The group has developed transgenic mouse models for NOR1, HOXA10, CK8 mutants, and multiple Sertoli cell-associated factors using transgenic overexpression and shRNA-mediated knockdown. These studies helped identify molecular regulators involved in spermatogenesis, male infertility, endometrial disorders, and cellular differentiation.

Collectively, the laboratory’s work has established versatile functional genomics platforms that bridge gene discovery with physiological and translational applications in animal biotechnology.