Multiple Strategies for Gene Transfer, Expression, Knockdown, and Chromatin Influence in Mammalian Cell Lines and Transgenic Animals

2006 ◽  
Vol 34 (3) ◽  
pp. 337-354 ◽  
Author(s):  
Félix Recillas-Targa
Keyword(s):  
Gene Transfer ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Transgenic Animals ◽  
Multiple Strategies ◽  
Mammalian Cell Lines

Applications of gene transfer to study rna splicing in mammalian cell lines

1993 ◽  
Vol 15 (2) ◽  
pp. 99-107
Author(s):  
Ian C. Eperon ◽  
Marion Hamshere
Keyword(s):  
Gene Transfer ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Rna Splicing ◽  
Mammalian Cell Lines

Gene transfer and expression in mammalian cell lines: A method to study regulation of neuropeptide processing

1989 ◽  
Vol 25 (7) ◽  
pp. A146
Author(s):  
James H. Meador-Woodruff ◽  
Benedetto Pellerito ◽  
Mary Hoversten ◽  
Donald Martin ◽  
Huda Akil
Keyword(s):  
Gene Transfer ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Mammalian Cell Lines

Caveolin-1 mediates gene transfer and cytotoxicity of polyethyleneimine in mammalian cell lines

2015 ◽  
Vol 402 (1-2) ◽  
pp. 203-211 ◽  
Author(s):  
Hai-Jie Yang ◽  
Pei Feng ◽  
Lei Wang ◽  
Zhi-Chao Li ◽  
Shuang-Ping Ma ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Caveolin 1 ◽  
Mammalian Cell Lines

scholarly journals Structure-guided selection of puromycin N-acetyltransferase mutants with enhanced selection stringency for deriving mammalian cell lines expressing recombinant proteins

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alessandro T. Caputo ◽  
Oliver M. Eder ◽  
Hana Bereznakova ◽  
Heleen Pothuis ◽  
Albert Ardevol ◽  
...  
Keyword(s):  
Cell Lines ◽  
Mammalian Cell ◽  
Recombinant Proteins ◽  
Cultured Cells ◽  
Hek293 Cells ◽  
Reaction Products ◽  
Enzyme Form ◽  
X Ray Crystallography ◽  
Mammalian Cell Lines ◽  
Apparent Loss

AbstractPuromycin and the Streptomyces alboniger-derived puromycin N-acetyltransferase (PAC) enzyme form a commonly used system for selecting stably transfected cultured cells. The crystal structure of PAC has been solved using X-ray crystallography, revealing it to be a member of the GCN5-related N-acetyltransferase (GNAT) family of acetyltransferases. Based on structures in complex with acetyl-CoA or the reaction products CoA and acetylated puromycin, four classes of mutations in and around the catalytic site were designed and tested for activity. Single-residue mutations were identified that displayed a range of enzymatic activities, from complete ablation to enhanced activity relative to wild-type (WT) PAC. Cell pools of stably transfected HEK293 cells derived using two PAC mutants with attenuated activity, Y30F and A142D, were found to secrete up to three-fold higher levels of a soluble, recombinant target protein than corresponding pools derived with the WT enzyme. A third mutant, Y171F, appeared to stabilise the intracellular turnover of PAC, resulting in an apparent loss of selection stringency. Our results indicate that the structure-guided manipulation of PAC function can be utilised to enhance selection stringency for the derivation of mammalian cell lines secreting elevated levels of recombinant proteins.

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