Table 1 

Co-targeting w and a gene-of-interest.

ShapeNumber of G0 whose G1 offspring had eyes that were:
G0 Lig4 genotypesgRNA plasmidDonor plasmidsFertile G0 (n)All redwhite & redCoffee & red/whiteAll WhiteCoffee & whiteAll coffee
Lig4169 aarmi-1 & w-1armi & w8.9%10102102
(78 nM)(132 nM ea.)(25/281)EJ: 1/10EJ: 2/10EJ: 0/2EJ: 1/1EJ: 1/2
HR: 0/10HR: 0/10HR: 0/2HR: 1/1HR: 2/2
Lig4169armi-1 & w-1armi & w17%3271111
armi-2 & armi-3(33 nM ea.)(43/260)EJ: 2/7EJ: 0/1EJ: 0/1EJ: 1/1EJ: 1/1
(26 nM ea.)HR: 2/7HR: 0/1HR: 1/1HR: 0/1HR: 1/1
Lig4+armi-3 & armi-4armi & w11%5142004
w-1 & w-1(132 nM ea.)(25/230)EJ: 0/5EJ: 0/1EJ: 1/3
(26 nM ea.)HR: 0/5HR: 0/1HR: 0/3
Lig4+armi-3 & armi-4armi & w20%2753333
w-1 & w-4(132 nM ea.)(44/220)EJ: 0/13EJ: 0/2EJ: 0/3EJ: 0/3EJ: 1/3EJ: 0/3
(26 nM ea.)HR: 0/13HR: 0/2HR: 1/3HR: 0/3HR: 1/3HR: 1/3
Lig4+armi-5 & armi-6armi & w29%31209013
w-1 & w-1(132 nM ea.)(64/220)EJ: 0/9EJ: 0/10EJ: 3/9EJ: 0/1EJ: 1/2
(26 nM ea.)HR: 0/9HR: 0/10HR: 0/9HR: 0/1HR: 0/2
Lig4+armi-5 & armi-6armi & w17%71212142
w-1 & w-1(132 nM ea.)(38/220)EJ: 0/5EJ: 0/6EJ: 2/6EJ: 1/1EJ: 3/3EJ: 1/2
(26 nM ea.)HR: 0/5HR: 2/6HR: 0/6HR: 0/1HR: 0/3HR: 1/2
Lig4+zuc-1 & zuc-2zuc & w15%8108232
w-1 & w-1(132 nM ea.)(33/222)EJ: 1/7EJ: 4/6EJ: 6/7EJ: 2/2EJ: 2/3EJ: 1/2
(26 nM ea.)HR: 0/7HR: 1/6HR: 3/7HR: 2/2HR: 3/3HR: 1/2
  • sgRNA-expressing and HR donor plasmids were coinjected into Lig4169 or Lig4+, vas-Cas9 G0 embryos. Shapes identify the corresponding experiment in Figure 2. n, total number of G0 embryos injected, irrespective of fertility or survival. “Coffee & red/white” includes G0 with coffee- and red-eyed, or with coffee-, white-, and red-eyed G1 broods. EJ, broods yielding indels; HR, broods yielding homologous recombinants (plasmid integration and gene conversion); conversion tracts were analyzed only for gene conversion events (Figure 5 and Figure 6).

  • a Coinjected with 1.2 µM of NLS-Cas9 protein (PNA-Bio, Inc., Thousand Oaks, CA), which had no observable effect.