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Allen Cell Collection
AICS-0107
SNPNM_000257.4(MYH7):c.1816G>A(p.Val606Met)
Gene SymbolMYH7
Gene Namemyosin heavy chain 7
Parental LineAICS-0075 cl. 85 ACTN2
Clone NumberClone TypeReplicateGenotype
9MutantAV606M/WT
17MutantAV606M/WT
63ControlBWT/WT
Certificate of Analysis

Obtain AICS-0107

2 mutant clones1 isogenic controls

AICS-0107

MYH7 in WTC-mEGFP-ACTN2 (mono-allelic tag)
Representative media for all clones
Cell line media

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 17 (V606M/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.

crRNA Target Site:5’ CCTCTCAATGAGACTGTCGTGGG 3’
DNA Donor Sequence:
Mutant* 5’ AGAACAAGGATCCTCTCAATGAGACTGTCATGGGCTTGTATCAGAAGTCTTCCCTCAAG 3’
WT 5’ AGAACAAGGATCCTCTCAATGAGACTGTGGTGGGCTTGTATCAGAAGTCTTCCCTCAAG 3’
Cas9: TrueCut™ Cas9 Protein
F Primer for PCR/Sequencing:5’ TGCATGATGACCTCCCACAC 3’
R Primer for PCR/Sequencing:5’ TGCAGAGCTGACACAGTCTG 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.1816G>A(p.Val606Met)
diagram
Top: ACTN2 locus showing 3 ACTN2 isoforms; Bottom: Zoom in on mEGFP insertion site at ACTN2 C-terminus