For cell-based assays, recent reports highlight the drive towards primary cell types, endogenous target expression and label-free systems3,4,5,6.
The expression of Na+/Ca2+ exchangers (NCXs) protein as an endogenous target in these regions was also studied.
Artificial TFs require an effector domain that controls the frequency of transcription initiation at endogenous target genes.
FnCas9 is unable to access an endogenous target in a certain chromatin configuration, but the binding of SpdCas9 at proximal locations alters the local chromatin homeostasis and enables FnCas9 to access and cleave the otherwise inaccessible target.
Most importantly, we have demonstrated that treatment of cultured human cells with PNAs led to expression of an endogenous target gene.
Specific trans-splicing, results in the fusion of the endogenous target mRNA of SLCO1B3 and the coding sequence of the suicide gene, provided by the RTM.
Thus we have successfully constructed a reporter cell line by engineering a reporter gene under control of an endogenous target gene promoter.
In dissociated neurons and brain slices, FingRs generated against PSD-95 and Gephyrin did not affect the expression patterns of their endogenous target proteins or the number or strength of synapses.
By editing an endogenous target site, we demonstrate that 3′ pairing determines targeting by specific miRNA family members and that seed pairing is not always sufficient for functional target interactions.
Targeted introduction of a double-stranded break (DSB) using designer zinc finger nucleases (ZFNs) in mammalian cells greatly enhances gene targeting – homologous recombination (HR) at a chosen endogenous target gene, which otherwise is limited by low spontaneous rate of HR.
The ability to introduce any chemical probe to any endogenous target protein in its native environment, that is in cells and in vivo, is anticipated to provide various new exciting tools for biological and biomedical research.
