Skip to main content

Advertisement

Table 2 Characterization of animals for the generation of a Syt7 conditional allele

From: Application of long single-stranded DNA donors in genome editing: generation and validation of mouse mutants

Founder ID Allele type 1 Allele type 2 Allele type 3 Copy number F1 animal ID PCR and sequencing outcome Copy number Allele 1 Allele 2
Syt7-4 cKO 5’ NHEJ + 3’ loxP Deletionc 1.03 ± 0.07 4.1a Only WT allele amplified 1.08 ± 0.04 WT Deletionc
4.1b Only WT allele amplified 1.02 ± 0.04 WT Deletionc
4.1c Only WT allele amplified 1.09 ± 0.06 WT Deletionc
  4.1d Both loxP present 1.99 ± 0.08 WT cKO
Syt7-8 cKO Appears homozygousa Additional insertionb 2.78 ± 0.15 8.1a Only WT allele amplified 1.00 ± 0.07 WT Deletionc
8.1b Only WT allele amplified 1.93 ± 0.08 WT WT
8.1c Both loxP present 2.98 ± 0.09 WT cKO + Additional insertionb
8.1d Only WT allele amplified 2.93 ± 0.09 WT WT + Additional insertionb
8.1e Both loxP present 2.07 ± 0.07 WT cKO
8.1f Both loxP present 2.13 ± 0.07 WT cKO
8.1g Both loxP present 2.89 ± 0.15 WT cKO + Additional insertionb
8.1h Only WT allele amplified 2.90 ± 0.15 WT WT + Additional insertionb
  1. The table summarizes the results of screening of the F0 animals obtained for the generation of a conditional Syt7 allele and of the F1 animals produced from the mating of F0 positive animals to WT mice. Outcomes of PCR and Sanger sequencing characterization employing the Syt7-F1 and Syt-R1 primers external to the lssDNA donor and copy counting of the donor, where relevant, are shown. Sequencing data showing a correct conditional allele is shown in Additional file 3: Figure S2a
  2. aSecond legitimate repair or combined with large deletion, unclear at F0 stage
  3. bRevealed by copy number, on or off target
  4. cDeletion including at least one external genotyping primer site