#Insulators are a class of regulatory DNA elements that possess a common ability to protect genes from inappropriate signals coming from their surrounding environment. Incorporating #tDNAinsulators into your construct could help you achieve superior yield.
Mechanism of action:
Insulators have 2 main functions:
#Heterochromatin barrier activity, which inhibits spreading of heterochromatin into neighboring euchromatin, and
#Enhancer blocking activity, which hinders the interaction between enhancers and promoters when placed between them.
Applications:
Insertion of #tDNA elements at both flanking sites of the expression units can intensively increase the expression of transgenes as well as the #cloning efficiency of the transfected cell pools. In #mAb producing cells, analysis of Ab expression in culture supernatant of stable pools and clones showed upto 3.1 and 9-fold increase in specific productivity respectively.
Insertion of #tDNA elements showed higher number of transgene expressing clones during single-cell cloning.
#Stability of Ab production is susceptible to modifications of transgene integration site and changes to the epigenetic profile at promoter sites. But clones having #tDNA elements showed 8 weeks of stability owing to barrier effect at flanking sites of transgene. #tDNA with AT-rich intervening sequences provided prolonged barrier activity by reducing the possibility of silencing through methylation of #CpG islands.
tDNA integration also increased levels of HC/ LC mRNA in transfected cells.
Faster recovery of transfected cells were also shown with tDNA insulator integration.
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