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Series GSE231138 Query DataSets for GSE231138
Status Public on May 05, 2023
Title Mammalian Genomic Manipulation with Orthogonal Bxb1 DNA Recombinase Sites for the Functional Characterization of Protein Variants
Organism synthetic construct
Experiment type Other
Summary The Bxb1 bacteriophage serine DNA recombinase is an efficient tool for engineering recombinant DNA into the genomes of cultured cells. Generally, a single engineered “landing pad” site is introduced into the cell genome, permitting the integration of transgenic circuits or libraries of transgene variants. While sufficient for many studies, the extent of genetic manipulation possible with a single recombinase site is limiting, and insufficient for more complex cell-based assays for protein function. Here, we harnessed two orthogonal Bxb1 recombinase sites to enable new avenues for mammalian synthetic biology. By designing plasmids with two recombinase sites, we demonstrate that we can avoid genomic integration of undesirable bacterial DNA elements. We also created “double landing pad” cells simultaneously harboring two orthogonal Bxb1 recombinase sites. These cells allow transgenic protein variant libraries to be readily paired with assay-specific protein partners or biosensors, opening up new functional readouts for large-scale functional assays.
Overall design Genomically engineered HEK293T cells with amplicons derived from exogenous DNA that has recombined using the Bxb1 bacteriophage integrase and various complementary Bxb1 attP and attB sites
Contributor(s) Matreyek KA, Roelle SM
Citation(s) 37922210
Submission date Apr 28, 2023
Last update date Nov 04, 2023
Contact name Kenneth Matreyek
Organization name Case Western Reserve University
Department Pathology
Lab Matreyek
Street address 2103 Cornell Rd
City Cleveland
State/province OH
ZIP/Postal code 44106
Country USA
Platforms (1)
GPL17769 Illumina MiSeq (synthetic construct)
Samples (19)
GSM7251895 Excision_flanked_replicate1_primerA
GSM7251896 Excision_control_replicate1_primerA
GSM7251897 Excision_flanked_replicate1_primerB
BioProject PRJNA962992

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Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE231138_Flanked_excision_results.csv.gz 390 b (ftp)(http) CSV
GSE231138_GN_recombination_results.csv.gz 1.2 Kb (ftp)(http) CSV
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Raw data are available in SRA
Processed data are available on Series record

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