15. CRISPR/Cas9CRISPR/Cas9
Genomic DNA
Genomic DNA
Donor DNA with
point mutation
Deletion
Donor with Reporter
Cas9
Guide RNA
GFP
GFP
Gene of interest
attP
Gene of interest
inserted at selected
locus
Genomic DNA TARGATT™ attP
docking site at safe
harbor locus
27. ゲノム DNA 増幅
T7 エンドヌクレアーゼⅠ切断
細胞集団
wt wt MU
wt wt MU
TALEN/CRISPR 誘導性 NHEJ
TALEN/CRISPR
DSB
wt
挿入欠失変異
MU
ミスマッチ切断
Gel
wt MU
変性と
再アニーリング
電気泳動による確認
Detection test of M. fementans-infected K562 cells
Amount of genomic DNA used for each lane
Target
Lane
gDNA
Lane
gDNA
DNA marker 0 ng 100 mg 50 ng 25 ng 12.5 ng 6.3 ng 3.2 ng
7 8 9 10 11 12 13 14 15
M N 1 2 3 4 5 6
1.6 ng 800 pg 400 pg 200 pg 100 pg 50 pg 25 pg 12.5 pg 6.25 pg