Obtaining chicken primordial germ cells used for gene transfer: in vitro and in vivo results

Journal of Applied Genetics
L Chojnacka-PuchtaAndrzej Plucienniczak

Abstract

Recently, several attempts have been made to create a generation of transgenic chickens via chimeric intermediates produced by primordial germ cells (PGCs) transfer. This study aimed to compare the influences of different chicken PGCs isolated from circulating blood (bPGCs) or gonads (gPGCs), purification (ACK, Percoll or trypsin) and transfection methods (electroporation or lipofection) on the expression of transgenes in vitro and the migration of modified donor cells to the recipient gonads. The highest average frequency of pEGFP-N1 plasmid-transfected bPGCs (75.8%) was achieved with Percoll density gradient centrifugation and electroporation. After ammonium chloride-potassium (ACK) treatment and lipofection, in vitro transgene expression was only detected in 35.2% of bPGCs. Chimeric chickens were produced from these purified, transfected and cultured cells, and the transgene was detected in the gonads of 44 and 42% of the recipient embryos that had been injected with bPGCs and gPGCs, respectively. These data confirmed that the combination of PGC purification via Percoll centrifugation and electroporation was an effective method for producing transgenic chickens. Subsequently, we used this method with expression vectors for g...Continue Reading

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Citations

Apr 1, 2018·Reproduction in Domestic Animals = Zuchthygiene·E ŁukaszewiczM Bednarczyk
Jan 15, 2019·Journal of the Science of Food and Agriculture·Muhammad SaeedSun Chao
Sep 19, 2019·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Andrea SvoradováPeter Chrenek
Jan 12, 2020·Journal of Applied Genetics·Luiza Chojnacka-Puchta, Dorota Sawicka
Jun 1, 2021·Poultry Science·Marek BednarczykEwa Grochowska

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Methods Mentioned

BETA
transgenic
fluorescence-activated cell sorting
transfection
density gradient centrifugation
FACS
PCR
density
enzyme-linked immunosorbent assay

Software Mentioned

SAS

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