Since scientists produced the first cloned mammal Dolly the sheep in 1996 using the somatic cellnuclear transfer technique, 23 mammalian species have been successfully cloned, including cattle, cats, dogs, horses and rats. Using this technique for primates had never been successful and no pregnancy had lasted more than 80 days. The main difficulty was likely the proper programming of the transferred nuclei to support the growth of the embryo. Tetra, a female rhesus macaque, was created by a team led by Gerald Schatten of the Oregon National Primate Research Center using a different technique, called "embryo splitting". She is the first "cloned" primate by artificial twinning, which is a much less complex procedure than the DNA transfer used for the creation of Zhong Zhong and Hua Hua. In January 2019, scientists in China reported the creation of five identical cloned gene-edited monkeys, using the same cloning technique that was used with Zhong Zhong and Hua Hua, and the same gene-editing CRISPR-Cas9 technique allegedly used by He Jiankui in creating the first ever gene-modified human babies Lulu and Nana. The monkey clones were made in order to study several medical diseases.
Process
Zhong Zhong and Hua Hua were produced by scientists from the Institute of Neuroscience of the Chinese Academy of Sciences in Shanghai, led by Qiang Sun and Muming Poo. They extracted nuclei from the fibroblasts of an aborted fetal monkey and inserted them into egg cells that had had their own nuclei removed. The team used two enzymes to erase the epigenetic memory of the transferred nuclei of being somatic cells. This crucial reprogramming step allowed the researchers to overcome the main obstacle that had precluded the successful cloning of primates until now. They then placed 21 of these ova into surrogate mother monkeys, resulting in six pregnancies, two of which produced living animals. The monkeys were named Zhong Zhong and Hua Hua, a reference to Zhonghua. Although the success rate was still low, the methods could be improved to increase survival rate in the future. By comparison, the Scotland-based team that created Dolly the sheep in 1996 required 277 attempts and produced only one lamb. The scientists also attempted to clone macaques using nuclei from adult donors, which is much more difficult. They implanted 42 surrogates, resulting in 22 pregnancies, but there were still only two infant macaques, and they died soon after birth.