Successful gene disruption of non-essential <i>L</i>. <i>mexicana</i> PKAC1 (LmxM.34.4010) and PKAC2 (LmxM.34.3960) with rRNA-P stable protocol.

<p>(A) PKAC1 (LmxM.34.4010) and PKAC2 (LmxM.34.3960) genes are located in chromosome 34 and share large part of conserved sequences. (B) Strategy used by rRNA-P stable protocol to disrupt LmxM.34.3960 and LmxM.34.4010 genes. Use targeting PKAC1 as an example, a gRNA was designed to target the gene specific 5’ end coding sequence of PKAC1 gene, which was then disrupted with the bleomycin resistance gene donor. (C) PCR analysis showing both PKAC1 gene alleles were successfully disrupted. (D) PCR analysis showing both PKAC2 gene alleles were successfully disrupted.</p> <p>(PDF)</p>