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Is it possible for a gene disrupted by an antibiotic resistance...
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+ Dna
+ Secretion
+ Crispr
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Matthew Leib
Is it possible for a gene disrupted by an antibiotic resistance...
I've been using the CRISPR/Cas9 system to induce a double strand break in my gene of interest, and incorporating a hygromycin resistance cassette with 40 bp homologous sequence to the break site. This is intended to disrupt the gene function and allow for easy screening of successful transformants. I've verified the insertion of the hyg cassette by sequencing. The insertion point of the hyg cassette is directly downstream of a predicted secretion signal in my gene. I imagine the secretion signal will still be translated, resulting in the theoretical secretion of the gene/hyg cassette construct. Assuming that the hyg cassette sequence insertion doesn't result in a frameshift, could it be possible that the genetic region downstream of the hyg cassette could still be properly translated? If this is then cleaved from the hyg cassette, could it still be a functional protein? Am I missing something obvious? Thanks!
