crRNA Target Site: 5’ GTTCAAGCTGAAGCAGCGGGAGG 3’ |
DNA Donor Sequence: Mutant* 5’ TGGAAACATGAAGTTCAAGCTGAAGCAGCAGGAGGAGCAGGCGGAGCCAGACGGCACTG 3’ |
Cas9:TrueCut™ Cas9 Protein |
F Primer for PCR/ |
R Primer for PCR/ |
Red = PAM Site, Blue = Mutation
AICS-0105
SNPNM_000257.4(MYH7):c.1106G>A(p.Arg369Gln) |
Gene SymbolMYH7 |
Gene Namemyosin heavy chain 7 |
Parental LineAICS-0075 cl. 85 ACTN2 |
| Clone Number | Clone Type | Replicate | Genotype |
|---|---|---|---|
| 22 | Control | A | WT/WT |
| 30 | Mutant | A | R369Q/WT |
| 31 | Mutant | A | R369Q/R369Q |
| 32 | Mutant | A | R369Q/WT |
| 57 | Mutant | B | R369Q/WT |
| 89 | Control | B | WT/WT |
AICS-0105
MYH7 in WTC-mEGFP-ACTN2 (mono-allelic tag)
One vial of distribution lot was thawed (cells were treated with ROCK inhibitor for 24hrs post-thaw - refer to culture protocol). A four panel image of clone 57 (R369Q/wt) is shown here and is a representative image for all clones in the collection (except clone 89). Cultures were observed daily. Colonies were imaged one (D1) and four (D4) days post-thaw using a Leica microscope at 4x and 10x magnification. Scale bars are shown.
EditingDesign
GenomicCharacterization
StemCellCharacteristics
CRISPR-Cas9 methodology was used to introduce a single base pair mutation to MYH7, and mEGFP at C-terminus of ACTN2 as shown below.
