caa a22 4500 465820743 CHVBK 20180323112135.0 cr unu---uuuuu 170327e19900901xx s 000 0 eng 10.1007/BF01233193 doi (NATIONALLICENCE)springer-10.1007/BF01233193 Targeted gene replacement at the endogenous APRT locus in CHO cells [Elektronische Daten] [Gerald Adair, Rodney Nairn, John Wilson, Julia Scheerer, Katherine Brotherman] We demonstrate the feasibility of targeted gene replacement at an endogenous, chromosomal gene locus in cultured mammalian cells, employing a two-step strategy similar to an approach routinely used for genetic manipulation in yeast. Utilizing an APRT+ recombinant generated by targeted integration of plasmid sequences (including a functional copy of the gpt gene) at the CHO APRT locus, we have been able to select gpt− "pop-out” recombinants that have arisen by intrachromosomal recombination between APRT direct repeats at the targeted integration site. Reciprocal exchanges leading to "pop-out” of integrated plasmid/gpt gene sequences occur at a rate of ≈6.3×10−6 per cell generation. Depending on the site of crossover, such "pop-out” events result in either replacement or restoration of the original APRT target gene sequence. Plenum Publishing Corporation, 1990 Adair Gerald Science Park-Research Division, The University of Texas M.D. Anderson Cancer Center, 78957, Smithville, Texas aut Nairn Rodney Science Park-Research Division, The University of Texas M.D. Anderson Cancer Center, 78957, Smithville, Texas aut Wilson John Marrs McLean Department of Biochemistry, Baylor College of Medicine, 77030, Houston, Texas aut Scheerer Julia Science Park-Research Division, The University of Texas M.D. Anderson Cancer Center, 78957, Smithville, Texas aut Brotherman Katherine Science Park-Research Division, The University of Texas M.D. Anderson Cancer Center, 78957, Smithville, Texas aut Somatic Cell and Molecular Genetics Kluwer Academic Publishers-Plenum Publishers 16/5(1990-09-01), 437-441 0740-7750 16:5<437 1990 16 11188 https://doi.org/10.1007/BF01233193 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01233193 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Adair Gerald Science Park-Research Division, The University of Texas M.D. Anderson Cancer Center, 78957, Smithville, Texas aut NATIONALLICENCE 700 1- Nairn Rodney Science Park-Research Division, The University of Texas M.D. Anderson Cancer Center, 78957, Smithville, Texas aut NATIONALLICENCE 700 1- Wilson John Marrs McLean Department of Biochemistry, Baylor College of Medicine, 77030, Houston, Texas aut NATIONALLICENCE 700 1- Scheerer Julia Science Park-Research Division, The University of Texas M.D. Anderson Cancer Center, 78957, Smithville, Texas aut NATIONALLICENCE 700 1- Brotherman Katherine Science Park-Research Division, The University of Texas M.D. Anderson Cancer Center, 78957, Smithville, Texas aut NATIONALLICENCE 773 0- Somatic Cell and Molecular Genetics Kluwer Academic Publishers-Plenum Publishers 16/5(1990-09-01), 437-441 0740-7750 16:5<437 1990 16 11188 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer