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AVA, LLC
DRAP Method
Step 1:
Pronuclear Injection
X
rDRAP + Mutant Oligonucleotide(s)
X
Gene of Interest
Exon Target
Fertilized Egg
1
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AVA, LLC
DRAP Method
Step 2:
Implantation
Mice born in ~ 21 days.
Implantation & Gestation
Fertilized Egg
Piglets born in ~114 days.
2
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AVA, LLC
DRAP Efficiencies In Mice
Gene & Target Characteristics
Genes
Strain
N-Myc
129
b1
Globin
129
c-kit
129
c-kit
DBF1
Natural Mutant
or
ES (- / -) KO
73%
Perinatal Lethal
Runts
DRAP
Oligo Strand
None
Sense
5’-OH
16
Normal
Yes
Sense
5’-OH
15
11 Stillborn
None
Sense
5’-OH
7
Normal
Yes
Sense
5’-OH
15
5 Runts With
Crenated RBCs
33%
Yes
Sense
5’-P
60
1 White-Spotted
2%
Sense
Sense
Antisense
Duplex - 5’ Overhang
Duplex - 3’ Overhang
5’-OH
5’-P
5’-OH
5’-OH
5’-OH
72
86
86
91
162
23 PL; 3 DC + LP
15 PL; 2 DC + LP
36 PL; 4 DC + LP
21 PL; 7 DC + LP
18 PL; 5 DC + LP
Yes
n
Observed
Phenotypes
Phenotype
Conversion
Oligo
Ends
3
14-50%
Perinatal Loss (PL)
White-Spotted
Dilute Coat Color (DC)
Hematologic Changes
[Lymphopenia (LP)]
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