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Targeted histone demthylation improves somatic cell reprogramming into cloned blastocysts_Meng and Oback.pdf (782.54 kB)

Targeted histone demethylation improves somatic cell reprogramming into cloned blastocysts but not postimplantation bovine concepti

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journal contribution
posted on 2023-05-03, 21:49 authored by Fan Li MengFan Li Meng, Kathrin Stamms, Romina Bennewitz, Andria Green, Fleur Oback, Pavla Turner, Jingwei WeiJingwei Wei, Björn ObackBjörn Oback
Correct reprogramming of epigenetic marks in the donor nuclei is a prerequisite for successful cloning by somatic cell transfer. In several mammalian species, repressive histone lysine methylation marks, in particular H3K9me3, form a major barrier to somatic cell reprogramming into pluripotency and totipotency. We engineered bovine embryonic fibroblasts for the doxycycline-inducible expression of Kdm4b, a demethylase that removes histone 3 lysine 9 trimethylation (H3K9me3) and H3K36me3 marks. Upon inducing Kdm4b, H3K9me3 and H3K36me3 levels reduced ~3-fold and ~5-fold, respectively, compared to non-induced controls. Donor cell quiescence has been previously associated with reduced somatic trimethylation levels and increased cloning efficiency in bovine. By simultaneously inducing Kdm4b expression (via doxycycline) and quiescence (via serum starvation), global H3K9me3 and H3K36me3 levels were further reduced by an average ~12-fold and ~38-fold, respectively, compared to non-induced, non-starved control fibroblasts. Following somatic cell transfer and sustained doxycycline induction during embryo culture, Kdm4b-BEF fibroblasts reprogrammed significantly better into cloned blastocysts than non-induced donor cells. However, detrimethylated donors and sustained Kdm4b-induction did not increase rates of post-blastocyst development from implantation to survival into adulthood. In summary, KDM4B only improved somatic cell reprogramming into early preimplantation stages, highlighting the need for alternative experimental approaches to reliably improve somatic cloning efficiency in cattle.

History

Rights statement

© The Author(s) 2020. Published by Oxford University Press on behalf of Society for the Study of Reproduction.

Publication date

2020-04-21

Project number

  • Non revenue

Language

  • English

Does this contain Māori information or data?

  • No

Publisher

Oxford University Press

Journal title

Biology of Reproduction

ISSN

0006-3363

Citation

Meng, F. L., Stamms, K., Bennewitz, R., Green, A., Oback, F., Turner, P., … Oback, B. (2020). Targeted histone demethylation improves somatic cell reprogramming into cloned blastocysts but not postimplantation bovine concepti. Biology of Reproduction, 103(1), 114–125. doi:10.1093/biolre/ioaa053

Funder

Ministry of Business Innovation & Employment

Contract number

A25101

Job code

15240