Variants of a gene using OriGene’s pre-designed donor vector
Posted: Fri 3 Mar 2017 10:21How to knockout all the splicing variants of a gene using OriGene’s pre-designed donor vectors, eg. OriGene’s CRISPR knockout / knockin kit?
FAQ from OriGene
Different splice variants of a gene are generated from the same pre-mRNA, spliced differently. When we design target sequences to knockout all the splicing forms of a gene, the target sequences are located around the start codon, ATG, of the longest splice variant. The 3’ end of the left homologous arm in the donor vector is right upstream of the start codon ATG. After homologous recombination, the GFP-puro cassette replaces part of the coding region of the longest splice variant in the genome. At the 3’ end of the GFP-puro functional cassette, there is a transcriptional stop signal; therefore the rest of the target gene will not be transcribed.
Access to all Origenes' vectors
FAQ from OriGene
Different splice variants of a gene are generated from the same pre-mRNA, spliced differently. When we design target sequences to knockout all the splicing forms of a gene, the target sequences are located around the start codon, ATG, of the longest splice variant. The 3’ end of the left homologous arm in the donor vector is right upstream of the start codon ATG. After homologous recombination, the GFP-puro cassette replaces part of the coding region of the longest splice variant in the genome. At the 3’ end of the GFP-puro functional cassette, there is a transcriptional stop signal; therefore the rest of the target gene will not be transcribed.
Access to all Origenes' vectors