Production of rhesus monkey cloned embryos expressing monomeric red fluorescent protein by interspecies somatic cell nuclear transfer.
Interspecies somatic cell nuclear transfer (iSCNT) is a promising method to clone endangered animals from which oocytes are difficult to obtain. Monomeric red fluorescent protein 1 (mRFP1) is an excellent selection marker for transgenically modified cloned embryos during somatic cell nuclear transfer (SCNT). In this study, mRFP-expressing rhesus monkey cells or porcine cells were transferred into enucleated porcine oocytes to generate iSCNT and SCNT embryos, respectively. The development of these embryos was studied in vitro. The percentage of embryos that underwent cleavage did not significantly differ between iSCNT and SCNT embryos (P>0.05; 71.53% vs. 80.30%). However, significantly fewer iSCNT embryos than SCNT embryos reached the blastocyst stage (2.04% vs. 10.19%, P<0.05). Valproic acid was used in an attempt to increase the percentage of iSCNT embryos that developed to the blastocyst stage. However, the percentages of embryos that underwent cleavage and reached the blastocyst stage were similar between untreated iSCNT embryos and iSCNT embryos treated with 2mM valproic acid for 24h (72.12% vs. 70.83% and 2.67% vs. 2.35%, respectively). These data suggest that porcine-rhesus monkey interspecies embryos can be generated that efficiently express mRFP1. However, a significantly lower proportion of iSCNT embryos than SCNT embryos reach the blastocyst stage. Valproic acid does not increase the percentage of porcine-rhesus monkey iSCNT embryos that reach the blastocyst stage. The mechanisms underling nuclear reprogramming and epigenetic modifications in iSCNT need to be investigated further.
Department of Animal Science, Agricultural College of Yanbian University, Yanji 133002, PR China.
Hai-Ying
HY
Zhu
Department of Animal Science, Agricultural College of Yanbian University, Yanji 133002, PR China.
Jin-Dan
JD
Kang
Department of Animal Science, Agricultural College of Yanbian University, Yanji 133002, PR China.
Suo
S
Li
Department of Animal Science, Agricultural College of Yanbian University, Yanji 133002, PR China.
Jun-Xue
JX
Jin
Department of Animal Science, Agricultural College of Yanbian University, Yanji 133002, PR China.
Yu
Y
Hong
Department of Animal Science, Agricultural College of Yanbian University, Yanji 133002, PR China.
Long
L
Jin
Department of Animal Science, Agricultural College of Yanbian University, Yanji 133002, PR China.
Qing
Q
Guo
Department of Animal Science, Agricultural College of Yanbian University, Yanji 133002, PR China.
Qing-Shan
QS
Gao
Department of Animal Science, Agricultural College of Yanbian University, Yanji 133002, PR China.
Chang-Guo
CG
Yan
Department of Animal Science, Agricultural College of Yanbian University, Yanji 133002, PR China. Electronic address: yinxj33@msn.com.
Xi-Jun
XJ
Yin
D000107:Acetylation / Q000187:drug effects; D000818:Animals; D019976:Cloning, Organism / Q000379:methods*; D018274:Electroporation / Q000379:methods; D015870:Gene Expression; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006655:Histone Deacetylases / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics*; D008253:Macaca mulatta / Q000196:embryology* / Q000235:genetics*; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D014635:Valproic Acid / Q000494:pharmacology; D000097573:Red Fluorescent Protein
D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases; D006657:Histones; D008164:Luminescent Proteins; D014635:Valproic Acid; D000097573:Red Fluorescent Protein
Electroporation; Histone acetylation; Rhesus monkey cell; VPA; iSCNT; mRFP1