Since 1997 when the first cloned mammal was born in Europe, there have been many abortive bids to use the technology in India. It was only in 2009, when Kashmir scientist, Dr Riyaz A Shah’s specially designed technique led to two cloned buffaloes in NDRI Karnal. Back home, after his PhD, he gave Kashmir the first cloned Pashmina goat, Noori, who is already a granny. In a detailed interview with Masood Hussain, Dr Riyaz explains his challenges and successes and his current research focus at SKUAST-K

KASHMIR LIFE (KL):  What is cloning and what are its applications?

DR RIYAZ AHMAD SHAH (DRAS): In normal conditions, animal breeding takes place by sexual reproduction, in which males and females physically get together to reproduce. However, cloning is an assisted reproductive technology, where the cells of either a male or female animal are taken and developed in laboratory conditions until an embryo is formed. It is then implanted in a surrogate mother. The offspring is born after it completes its gestation period. The process is efficient as it allows farmers to increase the number of their herds by providing more copies of the best-quality breed in the herd. In 1997, the world witnessed its first cloned mammal in the form of a sheep called Dolly, a female Finnish Dorset sheep cloned from an adult somatic cell

KL: Before we talk about your contributions to cloning, kindly tell us about your learning curve and the entire journey from your schools to SKUAST-K.

DRAS: I was born and raised in Batmaloo Srinagar. I did my early schooling at a local school and then joined Tyndale Biscoe for further studies. I aspired to be a doctor but couldn’t crack the entrance test; so, I ended up in veterinary science. I graduated from the Sher-e-Kashmir University of Agricultural Sciences and Technology (SKUAST). Owing to a good number of vacancies in the field, I got a job immediately after completing my degree. However, the thirst for learning more and being mentored by professors who had completed their studies from other states inspired me to go for further studies outside Kashmir. I cracked the prestigious national veterinary entrance test and was post-graduated from Indian Veterinary Research Institute Bareilly, Uttar Pradesh.

I came back to Kashmir and worked in the Department of Animal Husbandry for some years. In 1998, I joined SKUAST as an Assistant Professor. Initially, I was posted at Cattle Farm in Manasbal, Ganderbal. It proved to be a good learning experience. In 2005, I got admission as an in-service PhD candidate at National Dairy Research Institute. There, I came across a group who were working on cloning at that time. Interested, I joined them. The group had been working on a project of cloning buffalo. The group was struggling to form a cloned embryo since 1997 but could not succeed. I took the challenge and my PhD guide Dr S K Singla encouraged me for it. It took me nearly two years to standardize various techniques related to cloning but I succeeded.

KL:  What were the major takeaways of your PhD programme?

DRAS: The topic of my research was the production of handmade cloned embryos in buffalos. The embryo formed in the laboratory was transferred to a surrogate mother. It was sheer luck that I got the best quality cloned embryos. After completing the gestation period, a healthy buffalo was born on February 6, 2009, at National Dairy Research Institute (NDRI), Karnal. It was named Samrupa, the world’s first cloned buffalo calf. It made headlines throughout the world. I did not anticipate such a positive outcome. The calf, however, succumbed to lung infection a few days after it was born.

It was followed by another healthy and normal cloned calf named Garima, born on June 6, 2009.

KL: How many scientists were successful in the process of cloning development before you?

DRAS: My guide, Dr SK Singla, already had his PhD in clone generation under his credit but he remained unsuccessful in the formation of a live and healthy cloned progeny. During the course of my research, two other students were working on the same topic. However, they failed to get any positive results. Samrupa was the first live birth of a cloned buffalo at the institute and proved to be a milestone. Since then the institute has produced 20-25 cloned buffalos. The process involved in Samrupa and Garima acted as a road map for the researchers, who are now merging it with the science of gene editing to incorporate the selective qualities in the cloned organisms.

KL: When you were back home, you cloned Noori, the first Pashmina goat in March 2012?

DRAS: After I finished my PhD and returned to SKUAST, we started working on the Pashmina goat clone. We had to first set up facilities here at the SKUAST campus in Shuhama because we lacked the infrastructure. With project funding of Rs 2.50 crore from the Indian Council for Research (ICR), we were able to acquire basic equipment for our research.

Our objective was to develop a cloned embryo, implant it into a female and get a viable cloned organism.  Noori was one of the clones. While earlier researchers had tried to develop clones of various species but Pashmina goat was never experimented on. So, we had to start from scratch. We isolated and cultured the cells of the Pashmina goat. We conducted a study on the different species that can provide oocytes. We had to employ the Pashmina goat’s somatic cells and an egg from a different species.

Since people do not prefer goat meat in Kashmir, we had to get access to the ovaries of goats from a slaughterhouse in Delhi. This made the process a bit hectic and it took us two years to standardize the techniques. However, we got successful in the development of cloned embryos, which were then implanted into a surrogate mother. After 20-25 unsuccessful trials, Noori was our first live cloned Pashmina goat. Noori is currently living a normal and healthy life. It has given birth to 5-6 offspring via the natural reproductive process. Noori has also been a source of Pashmina wool like other naturally produced Pashmina goats.

Dr Riyaz A Shah (SKUAST-K)

KL: What are the differences between naturally reproduced organisms and cloned ones?

DRAS: A clone is genetically as good as a naturally bred organism. Cloning allows choosing the characters and traits we want in an organism, thus allowing farmers to increase the overall quality of their breed. Cloning also enables the production of the desired gender of a species. Farmers for example prefer a cow over a bull, cloning helps them have as many cows as they desire.

KL:  How different is Noori from her mother and her own offspring?

DRAS: Noori’s mother was a naturally bred Pashmina goat. Its embryo was implanted in a surrogate mother, who also happened to have naturally reproduced. After Noori’s birth, we studied its physiology and other parameters and found it and its progeny as good as any other Pashmina goat. We concluded that it can be used for the same purposes as we do use a normal goat. Also, there is no restriction on it or its progeny being used as food.

KL:  What has been your research focus since you completed the landmark Noori’s project?

DRAS: After Noori’s success, we approached various institutes for funding our projects. We got successful and secured a project, where we introduced gene editing in cloning. We tried to incorporate CRISPR-Cas9, a naturally occurring genome editing system in our research. We identified the gene responsible for Pashmina production in Noori and now we are trying to edit the gene so that the cloned progeny will be a source of good quality and improved quantity of Pashmina. We are nearly successful in creating gene-edited and cloned embryos. We are hopeful that we will soon be able to witness its progeny as well.

Simultaneously we are working on gene editing in sheep, where our focus is to increase meat production. This is a collaborative project with ICR, while scientists outside with the same objective are working on buffaloes; we are at the same time working on sheep.

KL: What ethical issues do genetically modified organisms (GMOs) face?

DRAS: One of the main ethical issues that GMOs face is their uncontrolled use. Many countries have allowed using GMOs as food. But yes scientists first have to make sure that gene editing does not lead to any abnormality in the organism.

(Humaira Nabi processed the interview)

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