Cas9-expressing cattle using the PiggyBac transposon all-in-one system

BMC Genomics

Table 1 Overview of microinjections and fluorescence-positive embryo transfer for Cas9-expressing cattle production

Group	No. injected oocyte	No. Blastocyst	No. RFP or GFP Blastocyst	No. recipient	No. offspring
PB-Cas9-RFP-FatI	794	151	34	5	4
PB-Cas9-GFP-sgPRNP	424	84	31	18	7

From the microinjection of 794 oocytes with the PB-Cas9-RFP-FatI vector, 151 blastocysts were developed and 34 exhibited RFP expression. A single RFP-positive blastocyst was transferred into each of five recipient animals, resulting in the birth of four calves. In the other group, the PB-Cas9-GFP-sgPRNP vector was microinjected alongside a transposase vector into 424 zygotes, yielding 84 blastocysts, 31 of which expressed GFP. One GFP-expressing blastocyst was transferred into each of 18 recipients, leading to seven confirmed pregnancies. Five pregnancies resulted in normal births, while two (#G1 and #G6) resulted in stillbirths due to dystocia

ISSN: 1471-2164