rna-i insulin-like androgenic hormones and monosex culture of prawns: the first implementation of...
TRANSCRIPT
![Page 1: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/1.jpg)
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
![Page 2: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/2.jpg)
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
&
![Page 3: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/3.jpg)
♂♀ 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
![Page 4: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/4.jpg)
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
![Page 5: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/5.jpg)
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.
![Page 6: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/6.jpg)
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
![Page 7: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/7.jpg)
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
![Page 8: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/8.jpg)
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 (%
)
![Page 9: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/9.jpg)
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
![Page 10: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/10.jpg)
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
![Page 11: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/11.jpg)
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.
![Page 12: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/12.jpg)
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
![Page 13: RNA-i Insulin-like androgenic hormones and monosex culture of prawns: The first implementation of RNA-interference in aquaculture Amir Sagi Ben-Gurion](https://reader031.vdocuments.site/reader031/viewer/2022033100/5697bfeb1a28abf838cb809a/html5/thumbnails/13.jpg)
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.