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Table 1 Comparison of integrase-based and nuclease-based approaches to engineer the Y-chromosome of Anopheles gambiae

From: Molecular tools and genetic markers for the generation of transgenic sexing strains in Anopheline mosquitoes

Donor plasmid Helper Plasmid Injected G0 embryos Surviving G0 larvae Transient G0 Larvae Transient G0 adult ♂ Pooled cages Transgenic G1s
A
 33 nM pHomeT 35 nM 1769 134 44 29 5 16/6559
ϕC31 integrase       1 pooled G0 transient cage
 33 nM pHomeT 35 nM 600 127 68 29 6 63/5147
attP-gRNA 3 pooled G0 transient cages
B
 40 nM attBCFP-VasaGFP 96 nM ~2000 n/a n/a 5 n/a 6/791
ϕC31 integrase
 52 nM 3xP3[AttP]RFP 90 nM ~4000 n/a n/a 15 n/a 11/6160
I-SceI
  1. (A) ΦC31-att recombinase was used to insert a vector, pHomeT, containing the attB donor sequence into the Y-attP strain, carrying the complementary attP target sequence on the Y-chromosome. CRISPR-cas9-directed knock-in was used to integrate the pHomeT plasmid into the same Y locus. (B) ΦC31-att recombinase was used to integrate a vector, attBCFPVasaGFP, containing the attB donor sequence into the Y-attP strain. I-SceI-directed knock-in was used to integrate the vector 3xP3[AttP]RFP into the Y chromosome of T4 strain, carrying the I-SceI recognition site [60].