Method for detecting a specific splice event of a gene of interest

Method according to any one of the preceding claims, wherein the expression product of the C-terminal splicing region of the split intein comprises at its C-terminus an asparagine.

Method according to any one of the preceding claims, wherein the split intein—heterologous polynucleotide construct further contains at least one polynucleotide encoding for a hetero-dimerizing domain or a homo-dimerizing domain, preferably at least one PDZ-domain or at least one coiled-coil-domain, more preferably two coiled-coil-domains in an antiparallel configuration, for accelerating the specific splice event.

Method according to any one of the preceding claims, wherein the split intein—heterologous polynucleotide construct further contains at least one polynucleotide encoding for a hetero-dimerizing domain or a homo-dimerizing domain, preferably at least one PDZ-domain or at least one coiled-coil-domain, more preferably two coiled-coil-domains in an antiparallel configuration, for accelerating the self-excision of the expression product of the split intein—heterologous polynucleotide construct from the expression product of the specific splice product.

Method according to any one of the preceding claims, wherein the heterologous polynucleotide of the split intein—heterologous polynucleotide construct further contains a temporary selection marker for stable cell line generation.

Method according to any one of the preceding claims, wherein detecting the heterologous polynucleotide and/or the expression product of the heterologous polynucleotide is carried out by any method selected from the group consisting of high-throughput screening, western blotting, mass spectrometry, luciferase-assays, and longitudinal live-imaging, preferably bioluminescence imaging, fluorescence imaging, photoacoustic imaging, MRI and PET.

Method according to any one of the preceding claims, wherein the method further comprises (iii) quantification of an isoform population of the protein of interest encoded by the gene of interest.

Method according to any one of the preceding claims, wherein the heterologous polypeptide of the split intein—heterologous polynucleotide construct is an antibiotic resistance gene and wherein the method alternatively to step (ii) or additionally to step (ii) comprises detecting the antibiotic resistance of the cells of interest comprising the protein of interest encoded by the gene of interest.

Method according to any one of the preceding claims, wherein the method alternatively to step (ii) or additionally to step (ii) comprises the detection of an isoform dependent cell-surface marker.

Method according to any one of the preceding claims, wherein the method further comprises (iii) manipulation of the folding process of the protein of interest encoded by the gene of interest.

Method according to any one of the preceding claims, wherein the method further comprises (iii) manipulation of the kinetics of the splice event of the gene of interest, preferably wherein the kinetics of the specific splice event is manipulated due to step (ii).

Method according to any one of the preceding claims, wherein the method further comprises (iii) enrichment of cells comprising the protein of interest encoded by the gene of interest, preferably enrichment of cells comprising a specific isoform of the protein of interest.

Method according to any one of the preceding claims, wherein the method further comprises (iii) modification of the folding process of the protein of interest.

Method according to any one of the preceding claims, wherein the method further comprises (iii) quantification of the protein of interest encoded by the gene of interest or quantification of the exon of interest.

Method according to any one of the preceding claims, wherein the method further comprises (iii) identification of a regulator of the inclusion or exclusion of the exon of interest, preferably identification of a regulator of the inclusion or exclusion of the exon of interest of a pre-mRNA.

Method according to any one of the preceding claims, wherein the split intein is gp41-1 or NrdJ-1.