Methods and compositions for monitoring and enhancing early embryo development
Method for determining developmental fate of early embryos comprises measuring the expression level of a gene selected from the group consisting of CDKN1C, IGF2R, MAGEL2, MKRN3, NAP1L5, NDN, PEG3, PHLDA2, TSSC4, and UBE3A genes. Also disclosed is a method for improving pregnancy rate, wherein early embryos whose expression level of the MKRN3, NDN, PEG3, PHLDA2, TSSC4, or UBE3A gene is not increased, or the expression level of the CDKN1C, IGF2R, MAGEL2, or NAP1L5 gene is not decreased, are selected for planting into a suitable uterus for further development. Also disclosed are methods for increasing the likelihood of an early embryo to develop successfully into full-term pregnancy, wherein a suitable amount of siRNA corresponding to the PHLDA2 gene is injected into a fertilized egg which is in turn cultured further and planted into a suitable uterus.
1. A method of improving the likelihood of a bovine embryo to develop successfully, the method comprising i) obtaining a supply of fertilized bovine eggs, ii) micro-injecting a suitable dosage of 5′-phospate-modified PHLDA2 siRNA into the fertilized bovine eggs, wherein blastocyst rate is increased compared to non-injected or sham injected controls; and iii) cultivating the fertilized eggs until they are ready for planting into a uterus.
2. The method of claim 1 , wherein the 5′-phospate-modified PHLDA2 siRNA comprises a duplex of PHLDA2 antisense [Phos]AGUAGCACCGGGCUAUAUCdTdT (SEQ ID No. 1), and PHLDA2 Sense GAUAUAGCCCGGUGCUACUdTdT (SEQ ID No. 2).
3. The method of claim 2 , wherein about 100 μM of the 5′-phospate-modified PHLDA2 siRNA is injected per fertilized egg.