http://rdf.ncbi.nlm.nih.gov/pubchem/patent/WO-2020254357-A1

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filingDate 2020-06-17-04:00^^<http://www.w3.org/2001/XMLSchema#date>
inventor http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_c82a4fba88abf028409fe2deb39eb7ee
publicationDate 2020-12-24-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber WO-2020254357-A1
titleOfInvention Method for the generation of a protein expressing cell by targeted integration using cre mrna
abstract Herein is reported a method for producing a recombinant mammalian cell comprising a deoxyribonucleic acid encoding a polypeptide and secreting the polypeptide comprising the steps of a) providing a mammalian cell comprising an exogenous nucleotide sequence integrated at a single site within a locus of the genome of the mammalian cell, wherein the exogenous nucleotide sequence comprises a first and a second recombination recognition sequence flanking at least one first selection marker, and a third recombination recognition sequence located between the first and the second recombination recognition sequence, and all the recombination recognition sequences are different; b) introducing into the cell provided in a) a composition of two deoxyribonucleic acids comprising three different recombination recognition sequences and one to eight expression cassettes, wherein the first deoxyribonucleic acid comprises in 5'- to 3'-direction, a first recombination recognition sequence, one or more expression cassette(s), a 5'- terminal part of an expression cassette encoding one second selection marker, and a first copy of a third recombination recognition sequence, and the second deoxyribonucleic acid comprises in 5'- to 3'-direction a second copy of the third recombination recognition sequence, a 3'-terminal part of an expression cassette encoding the one second selection marker, one or more expression cassette(s), and a second recombination recognition sequence, wherein the first to third recombination recognition sequences of the first and second deoxyribonucleic acids are matching the first to third recombination recognition sequence on the integrated exogenous nucleotide sequence, wherein the 5'-terminal part and the 3'-terminal part of the expression cassette encoding the one second selection marker when taken together form a functional expression cassette of the one second selection marker; c) introducing Cre-recombinase mRNA, and d) selecting for cells expressing the second selection marker and secreting the polypeptide, thereby producing a recombinant mammalian cell comprising a deoxyribonucleic acid encoding the polypeptide and secreting the polypeptide, wherein the Cre-recombinase mRNA is the only source of Cre-recombinase in the method.
priorityDate 2019-06-19-04:00^^<http://www.w3.org/2001/XMLSchema#date>
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http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ4QN19
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ97R22
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID226446360
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP44338
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ1JKW9
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCC0MBY9
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP33802
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ2RZH4
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ5M5S6
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCA7FQF9
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP09250
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID1252518
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID54685767
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ20EU8
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID226395685
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID1441136
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID3235715
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCC0MD73
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID61575096
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID58017467
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCB5QVE4
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID41711945
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ9HPQ9
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ9QNF7
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID34796
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCA7MMW6
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID48312
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP07605
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID54122
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCA8F7B3
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ0TNY3
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCB5ZCA9
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ5LHP5
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID17869
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID56418789
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID7987702
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ5WYR3
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ65WL7
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ9PQC8
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID59154857
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ9CKN6
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID29570061
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP63623
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID3239035
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP00931
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP63624
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID32100
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCB2TUN1
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ2SSX9
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ7VKJ0
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP63626
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID8627024
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCC5BDG9
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID45024431
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCB1IWZ4
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCC3P292
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID50018735
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ66624

Total number of triples: 670.