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Insulin-like androgenic hormones and monosex culture of prawns:
The first implementation of RNA-interference in aquaculture
Amir SagiBen-Gurion University of the Negev, Israel
2cm45 60 75 88 102 115 130 142 152
0
200
400
600
800
1000
Time (days)
Cu
mu
lati
ve W
eigh
t (g
/2m
2)
Modified from Sagi et al., 1986 Aquacult.
M. rosenbergii dimorphic growth pattern in favor of males
* Includes only the marketable yields (≥30g)
~60% higher incomeField study in IndiaNair et al., 2006
&
♂♀ Neo fe
mal
e
F1 (neo-female progeny) n.c.
F.S.
P.C.
Contr
ol
fem
ale
♂♀ F1 (control female progeny) n.c.
1 cm
3 mm
Ventura et al., Heredity, 2011
Mr-IAG silencing induces full functional sex reversal
Female specific sex marker
ZZWZ X
Neo ZZ
RNAi IAG
ZZ X
50% WZ
50%ZZ
100% ZZ
IAG silencing and prawn monosex biotechnologial cascade
Modified from Aflalo et al., 2006 Aquacult
The androgenic gland
Sex differentiation
Insulin-like
Biotechnology
Monosex all-male culture
Patent: RNA silencing of insulin-like gene in prawn and uses thereof. PCT/IL2009/000127 (priority: February 4, 2008) WO2009/098683, US2012/073508.
CA AA AA A AU CCC C GGG U U U UU U U3-’ -5’
AA CC GU GA
A A A A A A A A A A A A ACA AAAAA A AU C CC C C C GGG GU U U U U U UA CU U5-’ -3’
A T G C A A A A A A A A AT T T T T T T T TC C C C C CG G G G G5-’ -3’
A A A A A A A A AAT T T T T T T T T TC C C C C C CG G G G G G3-’ -5’DNA
RNA
How is RNAi working (non GMO)
Temporal dsRNA effect
Mr-IAGInsulin-like
androgenic hormone
A AUCCAU
AU C CG UU
AC G UC G
A AC CGU U
A AA C GU U U
AAA CGU U
AA GUU U
CA AC GU U
G
A gene encoding theInsulin like androgenic hormone
3 days
1 4 5 6 7 82 3T0 T0 T1 T1
Controls
PC NC
7 days
9 10 11 12 13 1514 16 17 18 19 20 21 2322 24
14 days
Clearance of exogenous dsRNA in injected prawns
Time (Days)3 7 14
RNAi temporarily reducesMr-IAG expression
Real time RT-PCR
Control 3 21 280
10
20
30
40
50
60
70
80
90
100
Days after dsMr-IAG injection
Rel
ativ
e M
r-IA
G e
xpre
ssio
n (%
)
Neo females (sex reversed males) reproductive performance
Females Neo females
Body weight (g) 32.03 ± 0.28 41.31 ± 0.22
BSI (%) 13.3 ± 1.63 12.7 ± 1.28
Eggs/g BW 859 ± 120 845 ± 67
Selective breeding scheme combined with all-male selective production
BGU line
1st Generation
Local strain
G1 G1
All-male PLs to growers
G2G2
BGU lineNeo
female
SelectedLocal strain
2nd Generation
All-male PLs to growers
BGU lineNeo
female
SelectedLocal strain
Modified from Aflalo et al., 2012
Recommendations• Protect from decay by crossing BGU with
Local lines (do not reuse crosses, no backflow of genes).
• Insert a selectively bred line into the portfolio (improved lines in Vietnam, India, or other sources).
• Pathogen free declaration of the BGU line.
Temporal RNAiNo hormone treatmentNo chemicalsNon GMO
Aquaculture production
enhancement
Up to this point, the technology
17.8 B $ 5.4 B $
EnvironmentalSolutions
Bio-control
PriceDelivery
Thank youDr. Tomer Ventura Ohad Rosen
Dr. Simy Weill Dr. Ziv Roth
Dr. Eli D. Aflalo Yaara Lazar
Dr. Rivka Manor Omri Sharabi
Dr. Lilah Glazer
Funding sources: BARD TIRAN Shipping Ltd.