AICS-0109
SNPNM_000257.4(MYH7):c.1573G>A (p.Glu525Lys)
Gene SymbolMYH7
Gene Namemyosin heavy chain 7
Parental LineAICS-0075 cl. 85 ACTN2
Clone NumberClone TypeReplicateGenotype
55ControlBWT/WT
56MutantBE525K/WT
72ControlBWT/WT
93MutantBE525K/WT
Certificate of Analysis

Obtain AICS-0109

2 mutant clones2 isogenic controls

AICS-0109

MYH7 in WTC-mEGFP-ACTN2 (mono-allelic tag)
Representative images for all clones
main image

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 93 (E525K/wt) is shown here and is a representative image for all clones in the collection. 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.

cRNA Target Site:5’ AAAGAGGCACCTTCTCGATGAGG 3’
DNA Donor Sequence:
Mutant* 5’ TCAGGTGGTAAGGCCAAAGAGGCACCTTCTTGATGAGGTCAATGC AGGCCTGCAGGTCC 3’
Cas9:TrueCut™ Cas9 Protein
F Primer for PCR/Sequencing: 5’ GGACTGTGTGGTGACAGAGG 3’
R Primer for PCR/Sequencing: 5’ GTGTGGGAGGTCATCATGCA 3’
Red = PAM Site, Blue = Mutation
CRISPR-Cas9 methodology was used to introduce a single base pair mutation to MYH7, and mEGFP at C-terminus of ACTN2 as shown below.
CRISPR-Cas9 methodology was used to introduce a single base pair mutation to MYH7, and mEGFP at C-terminus of ACTN2 as shown below.
Top: MYH7 locus showing 1 MYH7 isoform; Bottom: Zoom in on mutation site at isoform NM_000257.4(MYH7):c.1573G>A (p.Glu525Lys)
diagram
Top: ACTN2 locus showing 3 ACTN2 isoforms; Bottom: Zoom in on mEGFP insertion site at ACTN2 C-terminus