title: Split-intein mediated re-assembly of genetically encoded Ca(2+) indicators.
abstract: While genetically encoded Ca(2+) indicators (GECIs) allow Ca(2+) imaging in model organisms, the gene expression is often under the control of a single promoter that may drive expression beyond, the cell types of interest. To enable more cell-type specific targeting, GECIs can be brought under the, control of the intersecting expression from two promoters. Here, we present the splitting and, reassembly of two representative GECIs (TN-XL and GCaMP2) mediated by the split intein from Nostoc, punctiforme (NpuDnaE). While the split TN-XL biosensor offered ratiometric Ca(2+) imaging, it had a, diminished Ca(2+) response relative to the native TN-XL biosensor. In contrast, the split GCaMP2, biosensor retained similar Ca(2+) response to the native GCaMP2. The split GCaMP2 biosensor was, further targeted to the pharyngeal muscles of Caenorhabditis elegans where Ca(2+) signals from feeding C. elegans, were imaged. Thus, we envision that increased cell-type targetability of GECIs is feasible with two, complementary promoters.
authors: Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada. stanleysc.wong@utoronto.ca
authors: Stanley S C
authors: SS
authors: Wong
authors: Ippei
authors: I
authors: Kotera
authors: Evan
authors: E
authors: Mills
authors: Hiroshi
authors: H
authors: Suzuki
authors: Kevin
authors: K
authors: Truong
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D002460:Cell Line; D004258:DNA Polymerase III / Q000378:metabolism; D006801:Humans; D007202:Indicators and Reagents; D047668:Inteins; D008322:Mammals; D015335:Molecular Probes / Q000378:metabolism*; D046937:Nostoc / Q000378:metabolism; D010614:Pharynx / Q000378:metabolism; D019154:Protein Splicing
chemical_list: D007202:Indicators and Reagents; D015335:Molecular Probes; C074904:DNA polymerase III, alpha subunit; D004258:DNA Polymerase III; D002118:Calcium