Generation of a recloned transgenic cat expressing red fluorescence protein.
Somatic cells from a first-generation red fluorescence protein transgenic cat (first RFP TG cat) were used to produce a recloned RFP transgenic cat (Re-RFP TG cat) (Felis catus) that systemically expressed RFP. A total of 281 RFP cloned embryos were transferred into 13 surrogate mothers (mean=21+/-7.7 embryos/recipient). One surrogate cat was diagnosed pregnant (7.7%) and delivered one live kitten. The presence of the RFP gene in the mRNA and genomic DNA of the Re-RFP TG cat was confirmed by polymerase chain reaction analyses, and red fluorescence was detected in its internal organs and placental tissue samples. Analysis of nine feline-specific microsatellite loci confirmed that the Re-RFP TG cat was genetically identical to the donor cat. To test whether results such as normality of offspring and a low cloning success were due to epigenetic modifications, global methylation of placenta from the two first cloned RFP TG cats (77.08% and 82.29%) and the Re-RFP TG cat (76.38%) were compared by bisulfite mutagenesis sequencing analysis. In conclusion, although cloning efficiency was low, we demonstrated the successful use of a cloned first RFP TG cat as a donor cat to produce a Re-RFP TG cat. These results may facilitate future developments in biomedical models for human therapeutic applications.
Cho
S J
SJ
Division of Applied Life Science (BK21), Graduate School of Gyeongsang National University, Jinju, South Korea.
Bang
J I
JI
Yu
X F
XF
Lee
Y S
YS
Kim
J H
JH
Jeon
J T
JT
Yee
S T
ST
Kong
I K
IK
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D002415:Cats / Q000235:genetics*; D019976:Cloning, Organism / Q000379:methods / Q000662:veterinary*; D004247:DNA / Q000032:analysis / Q000235:genetics; D019175:DNA Methylation; D004624:Embryo Transfer / Q000662:veterinary; D005260:Female; D015870:Gene Expression; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics*; D018895:Microsatellite Repeats; D053652:Nuclear Transfer Techniques; D010920:Placenta / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D000097573:Red Fluorescent Protein
D004247:DNA; D008164:Luminescent Proteins; D012333:RNA, Messenger; D000097573:Red Fluorescent Protein