What do I need to do to get biallelic knock-out ?
Posted: Wed 8 Feb 2017 20:18Do I get monoallelic knock-out or biallelic knock-out using the knock-out kit via CRISPR? What do I need to do to get biallelic knock-out?
FAQ from OriGEne
If you isolate single cell colonies, in some cells gene knock-out may occur only in one allele; in some cells gene knock-out may occur in both alleles. If you only have single allele knockout and you want to get both-allele knockout, you can order another donor vector containing a different mammalian selection marker, such as blastocidin or neomycin resistant marker. OriGene has both functional cassettes. You can do the knockout procedure again with the new donor vector to target the second allele (one allele is already targeted and replaced with GFP-puro cassette). Alternatively, you can use Cre to flox out the puro cassette from your edited cells and use the same donor vector from the knockout kit and do the knockout again to target the second allele.
Access to all Origenes' vectors
FAQ from OriGEne
If you isolate single cell colonies, in some cells gene knock-out may occur only in one allele; in some cells gene knock-out may occur in both alleles. If you only have single allele knockout and you want to get both-allele knockout, you can order another donor vector containing a different mammalian selection marker, such as blastocidin or neomycin resistant marker. OriGene has both functional cassettes. You can do the knockout procedure again with the new donor vector to target the second allele (one allele is already targeted and replaced with GFP-puro cassette). Alternatively, you can use Cre to flox out the puro cassette from your edited cells and use the same donor vector from the knockout kit and do the knockout again to target the second allele.
Access to all Origenes' vectors