Generation of Transgenic Pigs by Cytoplasmic Injection of piggyBac Transposase-Based pmGENIE-3 Plasmids

Supplemental Data

Supplemental Figures S1-S4 and Tables S1-S6.

Files in this Data Supplement:

  • Supplemental Figures S1-S4 -

    Supplemental Figure S1. (A) Schematic representation of pmGENIE-3-EGFP. The locations of PCR primers are indicated by arrows. (B) Location in pmGENIE-3-EGFP to which the pB gene start and 3?-TRE Southern blot probe was constructed. (C) Location to which the EGFP Southern blot probe was constructed in pmGENIE-3-EGFP.

    Supplemental Figure S2. (A) New born piglets under Bright field and Fluorescence lighting. Only one piglet demonstrated variegated EGFP expression. (B) EGFP expression of various dissected organs from a male transgenic piglet and a male wild type piglet euthanized at the age of one week.

    Supplemental Figure S3. Southern blot analysis for all born transgenic pigs, done with a probe to the EGFP gene found in the pmGENIE-3-EGFP transposon. Transgenic pig TG9 is omitted in this analysis.

    Supplemental Figure S4. Identification of transposon insertion sites from the four first born transgenic piglets. The pB 5?-TRE sites are flanked by the chromosomal sequences where the transposon inserted. The TTAA sequence on the border signifies transpositional integration of the transposon.

  • Supplemental Tables S1-S6 -

    Supplemental Table S1. Primers for PCR.

    Supplemental Table S2. Concentration effect of CTI delivered pmGENIE-3-EGFP plasmid on the positive rate of parthenogenetic embryos expressing EGFP.

    Supplemental Table S3. Effect of cytoplasmic co-injection of pmGENIE-3-EGFP and mPB plasmids on the positive rate of embryos expressing EGFP.

    Supplemental Table S4. Comparison of gene transfer efficiency between the pBt based plasmid pmGENIE-3-EGFP and the non-insertional pEGFP-N1 plasmid during CTI.

    Supplemental Table S5. Effect of CTI treatment time points on the positive rate of IVF embryos expressing EGFP when injected with pmGENIE-3-EGFP.

    Supplemental Table S6. Production of transgenic pigs by CTI co-injection of pmGENIE-3-EGFP and mPB plasmids into IVF derived embryos.