Skip to main content

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

Ï•C31 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

Ï•C31 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].