Simultaneous Inhibition of Histone Deacetylases and RNA Synthesis Enables Totipotency Reprogramming in Pig SCNT...

Area of Science:

• Reproductive biology
• Developmental biology
• Genetics

Background:

• Somatic cell nuclear transfer (SCNT) combined with genome editing is crucial for creating specialized swine lineages.
• Low efficiency in cell reprogramming remains a significant hurdle in SCNT-based cloning.
• Histone deacetylase inhibitors (HDACis) are used to improve chromatin reprogramming in SCNT embryos.

Purpose of the Study:

• To investigate the effects of inhibiting both histone deacetylases and RNA synthesis on pig SCNT embryo development.
• To determine if combined inhibition enhances cell reprogramming and subsequent embryo development.

Main Methods:

• SCNT embryos were treated with histone deacetylase inhibitors (HDACis) alone or in combination with RNA synthesis inhibitors.
• Pre- and post-implantation development of treated SCNT embryos was evaluated.
• Blastocyst formation, cell numbers, and gene expression related to embryo genome activation were analyzed.

Main Results:

• Inhibiting RNA synthesis for up to 40 hours did not impede blastocyst formation or cell count in porcine embryos.
• Combined HDACi and RNA synthesis inhibition in SCNT embryos led to increased cell numbers and gene expression for embryo genome activation.
• This dual inhibition promoted complete somatic cell reprogramming, resulting in normal fetal and full-term development of SCNT embryos.

Conclusions:

• Concomitant inhibition of histone deacetylases and RNA synthesis significantly improves SCNT efficiency in pigs.
• This strategy enhances somatic cell reprogramming and early embryonic development.
• The combined treatment offers a promising approach to advance genome editing and SCNT applications in swine.