Page 2 of 14

DATA REVIEW RECORD:

Active Ingredient: dsRNA transcript comprising a Dv49 inverted repeat sequence derived
from Diabrotica virgifera and the genetic material neces sary for its production in corn (via plasmid vector P V

- ZMIR10781 )
Product Name: Plant

- Incorporated Protectant MON 87410 and MON 87411 [ EPA Reg. No. 524

- EUP

- RNU ; OECD Unique ID MON

- 87411

- 9] Company Name: Monsanto Company
Decision Number: 464546
DP Barcode : 402217
Submission Number: NA

MRID No:
48919001 Molecular characterization Data for MON 87410 and MON 87411 Supporting an Experimental Use Permit

to Test MON 87410, MON 87411, Combined Trait Insect Control Corn, Combinations Necessary for their Production , and Controls

.
48919002 Assessment of CP4 EPSPS and Cry3Bb1 Protein Levels in Corn Tissues from MON 87410 a nd MON

87411 Produced in a United States Greenhouse trial during 2011

.
48919003 Assessment of DvS nf7 RNA levels in corn tissues from MON 87410 and MON 87411 Produced in a

United States Greenhouse trial during 2011

.
48919004 Preliminary Information to Characterize the Activity Spectrum of the dsRNA DvSnf7 Against a range of

Insect Taxa.



### B ACKGROUND

Monsanto Company is seeking a n Experimental Use Permit (EUP) to test a new plant

- incorporated protectant (PIP) technology, MON 87410 and MON 87411 , com bined trait insect control corn. Development and testing of new PIPs and PIP combinations providing multiple modes of action against corn rootworm (CRW) and Lepidopteran corn pest s is an important component of producing highly efficacious produc ts that als o support durable Insect Resistance Management (IRM)

. The PIP events comprising the combined trait produc ts in this EUP request include six registered corn events (MON 89034, EPA Reg. No. 524

- 576; TC1507, EPA Reg. No. 68467

- 2; MIR162, EPA Reg. No. 67979

- 1 4; MON 88017, EPA Reg. No. 524

- 551; and DAS

- 59122

- 7, EPA Reg. No. 68467

- 5; and EPA 5307, EPA Reg. No. 667979

- EU ) , and two corn events representing a new highly

- effective class of PIP against
Diabrotica spp . (MON 87410 and MON 87411).

Using biotechnology methods, Monsanto Company has developed MON 87410 and MON 87411, that confer resistance to corn rootworm (CRW; Diabrotica spp.). MON 87410 and MON 87411 contain a suppression cassette with an inverted repeat sequence (Dv49) derived from Diabrotica virgifera. The expression of the Dv49 suppression cassette results in the formation of a double stranded RNA (dsRNA) transcript. Upon consumption by the CRW pest, the plant

- produced Dv49 dsRNA in MON 87410 and
MON 87411 result in the down

- regul ation of the targeted Snf 7 (this gene is only in CRW) gene and eventual CRW mortality. MON 87410 and MO N 87411 also produce the Cry3Bb1 protein to protectPage 3 of 14

against CRW larval feeding. In addition, MON 87410 and MON 87411 are Roundup Ready ® corn events that produce the 5

- enolpyruvylshikimate

- 3

- phosphate synthase protein from Agrobacterium sp. strain CP4 (CP4 EPSPS), which confers tolerance to glyphosate, the active ingredient in Roundup 1 ® agricultural herbicides.

All of the above registered corn PIP products

, produce active ingredients derived from Bacillus thuringiensis (B.t.). Crystal proteins derived from B.t. are highly efficacious against corn pests, have a high

- degree of target specificity, and have been safely used in biotechnology

- de rived crops for over a
decade. The Cry proteins affect a mid

- gut pathology in susceptible insects by binding to receptors in the insect gut and causing ulceration. The Dv49 dsRNA component in MON 87410 and MON 87411 is currently being specifically evaluat ed by Monsanto for safety and efficacy. T he PIP events comprising the single and combined trait products in this request include o ther registered PIP Bacillus thuringiensis ( Bt ) proteins and either of the two new corn events MON 87410 and 87411. The Bt proteins to be used in the single or combination traits in the proposed EUP i nclude Cry1A.105, Cry2Ab2, Cry 1 F, Vip3Aa20, Cry3Bb1, Cry34Abl/Cry35Abl and eCry3.1Ab. The environmental and human health safety of these proteins has been demonstrated, and they are exempted from the requirement of a tolerance (40

CFR § 174.501, 174.502, 174.506, 174.518, 174.519, 174.520, 174.528, 174.532). A permanent tolerance exemption has been established for nucleic acids ( including the dsRNA ) that are part of the PIPs (40 CFR § 174.507). M arker proteins phosphinothricin acetyl transferase protein ( PAT ) , CP4 EPSPS, and phosphomannose isomerase protein ( PMI ) are also exempt from the requirement of a tolerance (40 CFR § 174.522, 174.523, 174.527).

Monsanto is applying for an e xperimental use permit (EUP) to conduct field testing and evaluate these single and combined events, corn PIP products that include two primary protocols sp anning two years from February 11 , 2013 through July 31, 2014

. A total of 20,893 acres are requested for this EUP across 22 U.S. states and Puerto Rico over two years. This include s 17,740 acres of PIP m aterials, 1,782

acres o f non

- PIP material, and 1,371 acres of border materials. The quantity of acres being requested
for 2013

- 2014 is require d to conduct field trials, using an extensive number of single and combined trait products, and generate data necessary to register a combined trait product under FIFRA.

The p roposed planting includes field testing the two corn events (MON 87410 and MON 87411) representing a new class of PIP s active against CRW ( Diabrotica spp

. ) with t he following combination PIP corn products: MON 89034 × MON 88017 [EPA Reg. No. 524

- 576], MON 89034 × TC1507 [EPA Reg. No. 524

- 585], TC1507 × DAS

- 59122

- 7[EPA Reg. No. 68467

- 6], MON 89034 × TC1507 ×MON
88017 × DAS

- 59122

- 7[EPA Reg. No. 524

- 581, 524

- 595], MON 89034 × TC1507 × MON 88017 [EPA Reg. No. 524

- 587], M ON 89034 × TC1507 × DAS

- 59122

- 7 [EPA Reg. No. 524

- 588], MON 88017 ×
DAS

- 59122

- 7[EPA Reg. No. 524

- 586] and MON 89034 × MON 88017 × DAS

- 59122

- 7 [EPA Reg. No.
524

- 589]. The PIP events comprising the combined trait products in this EUP include five registered corn events (MON 89034, EPA Reg. No. 524

- 576; TC1507, EPA Reg. No. 68467

- 2; MIR162, EPA Reg. No.67979

- 14; MON 88017, EPA Reg. No. 524

- 551; DAS

- 59122

- 7, EPA Reg. No.68467

- 5 and
5307 EPA Reg. No. 67979

- EE ). The following sections summarize the regulatory i nformation for each PIP corn event present in the combination PIP corn products:

Event MIR162 : The Vip3Aa20 protein produced in corn event MIR162 (EPA Reg. No. 67979

- 14) is derived from Bacillus thuringiensis protein Vip3Aa20, which is highly toxic to the major

1 ® Roundup and Roundup Ready are registered Trademark of Monsanto Technology LLC.Page 4 of 14

lepidopteran pests of corn, but is non

- toxic to humans, and was evaluated by EPA (U.S. EPA 2008) . The Vip3Aa20 protein does not exhibit meaningful similarity to known mammalian toxins or allergens, is readily digested in simulated gastric fluid a nd is not active after heat

treatment (U.S. EPA, 2009) . The EPA reviewed the safety of the Vip3Aa20 protein and granted an exemption from the requirement of a tolerance in corn in 2008 (40 CFR §174.501/528).

Event TC1507 : The Cry1F protein produced in eve nt TC1507 is a truncated form of the native
full

- length Cry1F protein from Bacillus thuringiensis var. aizawai . The Cry1F protein does not
exhibit sim ilarity to known mammalian toxins or allergens, is readily digested in simulated gastric fluid and is not active after heat treatment (U.S. EPA, 2010a). The Cry1F protein as produced in TC1507 was reviewed by EPA and granted an exemption from the requirement of a tolerance in corn in 2001, 2007 (40 CFR §174.520).

Event 5307 M aize was developed through Agrobacterium

- mediated transformation (via plasmid

vector PV

- ZMIR245) to express eCry3.1Ab protein for use as a plant

- incorporated protectant (PIP). This proposed PIP is a chimeric protein

, composed of portions of Bt Cry1Ab and modified Cry3A proteins and provides protection to corn from feeding damage caused by certain corn rootworm ( Diabrotica ) species. The Agency also established an exemption from the requirement of a tolerance for eCry3.1Ab protein residues when used as a PIP in corn [40 CFR § 174.532].

DAS 59122

- 7 : The Cry34Ab1 and Cry35Ab1 proteins produced in event DAS

- 59122

- 7 (EPA Reg. No.68467

- 5) form a binary insecticidal protein that protect s from damage by corn

rootworm ( Diabrotica spp. ). The amino acid sequences of the plant

- expressed Cry34Ab1 and Cry35Ab1 proteins are identical to the Cry34Ab1/Cry35Ab1 proteins produced by the host Bacillus thuringiensis strain PS149B1. The Cry34Ab1 and Cry35Ab1 proteins do not exhibit similarity to known mammalian toxins or allergens and are moderately or rapidly digested in simulated gastric fluid ( U.S. EPA, 2005b ). EPA reviewed the safety assessment of the Cry34Ab1 and Cry35Ab1 proteins as produced and granted an exemption from the requirement of a
tolerance in corn in 2005 (40 CFR §174.506).

Event MON 88017 : The Cry3Bb1 protein produced in MON 88017 (EPA Reg No. 524

- 551), is
a member of the Cry3Bb class of proteins that share >95% amino sequence homology. The Cry3Bb1protein is also produced in MON 87410 and MON 87411. The transformation plasmid (PV

- ZMIR10871), used to produce MON 87410 and MON 87411, contains expression cassette for Cry3Bb1 protein that has the identical amino acid sequences to the Cry3Bb1 produced in MON 88017. The expression and identity of the C ry3Bb1 pr otein from this cassette was

c onfirmed by SDS

- PAGE and determination of the N

- terminus to be identical to corn event
MON 86 . The Cry3Bb1 protein does not exhibit similarity to known mammalian toxins or allergens, is readily digested in simulated gastric fluid and is not immunodetectable in corn grain
after heat treatment (U.S. EPA, 2010b). The EPA previously reviewed the safety assessment of the Cry3Bb1 protein and granted an exemption from the requirement of a tolerance in corn in 2004 (40 CFR §174.518).

Event MON 89034 : The Cry2Ab2 protein produced in MON 89034 (EPA Reg. No. 524

- 575) is
>99% identical to the Cry2Ab2 protein produced in Bollgard II cotton (EPA Reg. No. 542

- 522). The Cry2Ab2 protein does not exhibit similarity to known mammalian toxins or allergens in
humans or animals, is readily digested in simulated gastric fluid and is not immunodetectable inPage 5 of 14

corn grain after heat treatment (U.S. EPA, 2008b). EPA reviewed safety assessment of the Cry2Ab2 protein as produced in MON 89034 and granted a n exemption from the requirement of a tolerance (40 CFR §174.503).

Event MON 89034 : Cry1A.105 produced in MON 89034 (EPA Reg. No. 524

- 575) is a Bt Cry1A

- type protein comprised of portions of naturally occurring Cry1Ab, Cry1Ac, and Cry1F proteins. As a result, Cry1A.105 shares 93.6% amino acid sequence identity with Cry1Ac, 90.0%
with Cry1Ab, and 76.7% with Cry1F proteins produced in insect

- protected corn and cotton. As with other Cry1A proteins, the Cry1A.105 protein does not exhibit similarity to know n mammalian toxins or allergens, is r eadily digested in simulated gastric fluid and is not immunodetectable in corn grain after heat treatment (U.S. EPA, 2008a). EPA reviewed the safety assessment of the Cry1A.105 protein as produced in MON 89034 and gran ted an exemption from the requirement of a tolerance in 2006 (40 CFR §174.502).

Corn events MON 88017, MON 87410 and MON 87411 produce the 5

- enolpyruvylshikimate

- 3

- phosphate synthase protein from Agrobacterium sp . Strain CP4 (CP4 EPSPS), which confers tolerance to glyphosate herbicide; corn events TC1507 and DAS

- 59122

- 7 produce the phosphinothricin acetyl transferase protein (PAT) from Streptomyces viridochromogenes which confers tolerance to glufosinate

- ammonium herbicide; and corn events MIR162 and 53 07 produce the phosphomannose isomerase protein (PMI) from Escherichia coli , which confers a laboratory positive selectable marker phenotype. Each of these three non

- pesticidal inert ingredients/ marker proteins have been thoroughly evaluated by EPA in as sociation with their respective PIP event registrations, and have been granted an exemption from the requirement of
a tolerance in corn.

To satisfy the data requirements specific for justifying an EUP for MON 87410 and MON 87411( Dv49 inverted rep eat sequ ence), Monsanto Company has submitted product characterization data which are reviewed in this report.

RECOMMENDATION:

T he product characterization and human health data submitted for Dv49 as expressed MON 87410 and

MON 87411 [ EPA Reg. No. 524

- EUP

- RNU ] are sufficient to support an Experimental Use Permit

.

Table 1 . Product Characterization Data Submitted for MON 87410 and MON 87411

STUDY TITLE STUDY SUMMARY MRID NO.

Molecular characterization
data for MON 87410 and MON 87411 Supporting an Experimental Use
Permit to Test MON 87410, MON 87411,
Combined Trait Insect Control Corn, Combinations Necessary Molecular characterization of MON 87410 and MON 87411 by
Southe rn blot analyses confirmed that the integrated DNA was inserted into the corn genome at a single locus in each event. Each event contained one copy of the DvSnf suppression cassette,
the cry3Bb1 and the cp4 epsps expression cassettes. No backbone DNA seque nces from PV

- ZMIR10871 were detected in either

MON 87410 or MON 87411. 48919001Page 6 of 14

STUDY TITLE STUDY SUMMARY MRID NO.

for their Production and
Control CLASSIFICATION: ACCEPTABLE

Assessment of CP4 EPSPS and Cry3Bb1 protein
levels in Corn tissues from
MON 87410 an d MON
87411 Produced in a
United States Greenhouse Trial during 2011 The concentration o f CP4 EPSPS and Cry3Bb1 protein were measured via ELISA in MON 87410 and MON 87411 corn events grown in a greenhouse trial. Expression of the Cry3Bb1 cassette resulted in production of the highest mean levels of Cry3Bb1 protein in the root (840 and 980 μg/g dry weight for MON 87410
and MON 87411, respectively). Production of Cry3Bb1 protein was lowest in the seed of each line (means of 4.1 and 4.4 μg/g dry weight for MON 87410 and 87411, respectively). Other tissues evaluated for production of Cry3Bb1 included leaf, forage, pollen
and silk and the mean levels of Cry3Bb1 ranged from 51 μg/g dry weight (MON 87410 pollen) to 400 μg/g dry weight (MON 87411 leaf). The mean CP4 EPSPS protein levels were highest in root (81 and 66 μg/g dry weight for MON 87410 and MON 87411, respectively) and lowest in seed (means of 3.6 and 3.0 μg/g dry weight in MON 87410 and MON 87411, respectively). Similarly, the same tissues were evaluated for production of CP4 EPSPS protein and ranged from a mean of 14 μg/g dry weight (MON 87411 forage) to 43 μg/g dry w eight in silk of both lines.

CLASSIFICATION: ACCEPTABLE 48919002

Assessment of Dv49
RNA Levels in Corn
Tissues from MON 87410
and MON 87411 Produced
in a United States Greenhouse Trial during
2011 MON 87410 and MON 87411 were grown in a U.S. greenhouse trial during 2011. Tissue samples were collected and analyzed for Dv49 level using a QuantiGene Plex 2.0 Assay. These data provide an estimation of the Dv49 level ( MON 87410: Leaf 0.0147 μg/g dwt; MON 87411: leaf 0.0132 μg/g dwt) on a fresh weight and dry weight basis in five tissue types throughout the growing season and in the starting seed.

CLASSIFICATION: ACCEPTABLE 48919003

Preliminary Information to characterize the Activity Spectrum of the dsRNA DVSnf7 Against a range
of Insect Taxa Efficacy against the target organism WCRW and the closely
related SCRW was demonstrated with larvae in 12

- day
concentration

- response diet bioassays. Adverse effects were not detected against the Chrysomelidae species, Leptinotarsa
decemlineata , or in representative species from the other three
families tested in the order Col eoptera at exposure c oncentrations that were reported to exceed field exposure levels. Additionally, no adverse effects were detected in representative species from the orders Hemiptera, Hymenoptera, or Lepidoptera. Results from bioassays demonstrated that the spectrum of activity of DvSnf7_240 dsRNA among the insects tested is restricted to the subfamily Galerucinae in the family Chrysomelidae within the order Coleoptera under the test conditions

.

CLASSIFICATION: SUPPLEMENTAL 48919004Page 7 of 14

Tolerance

This application is for a non

- food/feed use EUP. Tolerance exemptions exist for all active and inert ingredients produced by the PIP products under this EUP : eCry3.1Ab produced by 5307 corn: Dv49 dsRNA, Cry1A.105, Cry2Ab2, Cry1F, Vip3Aa20, Cry3Bb1, Cry34A b1/ Cry35Ab1, eCry3.1Ab, CP4 EPSPS, PAT, and PMI proteins.

Safety Assessment The majority of PIPs used within this EUP proposal consist of registered active ingredient Cry proteins
used in corn with widespread commercial corn production and highly successful safe and effective use s as PIPs in major crops within U.S. agriculture. These PIPs have been combined through breeding for
various assessments under this EU P. Two corn events within this proposed EUP are unregistered PI Ps.
MON 87410 and MON 874 11 produce the Cry3Bb1 protein present in the registered prod uct MON 88017, and also produce the CRW

- active Dv49 dsRNA that is designed, from the CRW target itself, to
deliver a very high level of t arget specificity . Since RNA suppression activity operates on a nucleic acid

sequence

- specific basis, the spectrum of activity is anticipated to be very narrow, and likely narrower than that observed for many B.t . proteins ( Whyard et al ., 2009 ).

P roduct Characterization

Monsanto Company has developed corn events MON 87410 and MON 87411 which express Dv49, Cry3Bb1 and CP4 EPSPS insecticidal proteins. MON 87410 and MON 87411 contain a suppression cassette with an inverted repeat sequence (Dv49) derived from Diabrotica virgifera . The expressio n of the Dv49 suppression cassette results in the formation of a double stranded RNA (dsRNA) transcript. Upon consumption, the plant

- produced Dv49 dsRNA in MON 87410 and MON 87411 is recognized by the CRW’s RNA interference (RNAi) machinery resulting in th e down

- regulation of the targeted Snf7 gene expression and eventual CRW mortality MON 87410 and MON 87411 also produce a modified Bacillus thuringiensis (subsp. kumamotoensis ) Cry3Bb1 protein to protect against CRW larval feeding. In addition, MON 87410 and MON 87411 produce the 5

- enolpyruvylshikimate

- 3

- phosphate synthase protein from Agrobacterium sp . Strain CP4 (CP4 EPSPS), which confers tolerance to glyphosate

. Cry3Bb1 is the same protein in the existing EPA registered PIP event MON 88017.

A. Transformation System
The Dv 49 suppression cassette and cry3Bb1 and cp4 epsps genes were transferred into the genome of corn cells by Agrobacterium tumefaciens mediated transformation utilizing the plant transformation

vector PV

- ZMIR10871. The transformation vector, PV

- ZMIR10871, used to develop MON 87410 and
MON 87411 is a vector that contains well

- characterized DNA segments required for selection and replicati on of the plasmid vector in bacteria and transfer of the T

- DNA into plant cells. The inserted T

DNA from this plasmid contains three gene cassettes.

1) The first cassette c ontain an inverted repeat sequence (named Dv49) derived from Diabrotica virgifera

, the Western Corn rootworm (WCR)

. The WCR Snf7 ortholog (DvSnf7) is a component of
the ESCORT

- III complex (endosomal sorting complex required for transport), which is involved in essential biological processes including sorting of cell membrane receptors ( Sweeney NT, et al

, 2006; Vaccari T, et al , 2009; Kim DW, et al , 2011). Due to its vital cellular function resulting in WCR mortality at relatively low concentrations when targeted by dsRNA, DvSnf7 was selected asPage 8 of 14

the target mRNA to investigate the molecul ar mechanisms leading to WCR mortality after feeding on dsRNA (Baum JA, et al , 2007). Once the dsRNA is ingested by WCR, it is thought to be taken up by midgut cells and processed by native RNAi machinery. The RNAi pathway is initiated by cleavage of d sRNA into short interfering si RNAs by the nuclease Dicer (Fire A, et al , 1998). The siRNAs then bind to a complex of protein know as RNA induced silencing complex (RISC) which leads to specific suppression of the target mRNA. The specific suppression can cause lethality if the target mRNA encodes a protein with an essential function within the insect.

DV49 dsRNA is 240 b ase pair (bp) . Dv49 siRNA i s 21

- 24 mer. High molecular weight (HMW)
northern blot shows all detecta ble Dv49 sequences are double stranded. Low molecular weight (LMW) northern blot showed siRNA production. The dsRNA with 240 bp is the active molecule that kills CRW in diet bioassay. Single stranded and small siRNA are not active against CRW in diet bioassays.

2) The second cas sette contains cry3Bb1 is a modified Bacillus thuringiensis (subsp. kumamotoensis ) protein to protect against corn rootworm larval feeding.

3) The third cassette produce s the 5

- enolpyruvylshikimate

- 3

- phosphate synthase protein from Agrobacterium sp. strain CP4 (CP4 EPSPS), which confers tolerance to glyphosate, the active ingredient in Roundup ® agricultural herbicides.

B. Protein Expression Enzyme

- linked immunosorbent assay (ELISA) analyses quantified the amount of Cry3B b1 and CP4
EPSPS protein concentrations expressed in leaf, root , forage, pollen, silk and seed plant samples
collected from PIP events MON 87410 and MON 87411. The results of the ELISA analysis showed expression of the Cry3Bb1 cassette resulted in production of the highes t mean levels of Cry3Bb1 protein in the root (840 and 980 μg/g dry weight for MON 87410 and MON 87411, respectively). Production of Cry3Bb1 protein was lowest in the seed of each line (means of 4.1 and 4.4 μg/g dry weight for MON 87410 and 87411, respecti vely). Other tissues evaluated for production of Cry3Bb1 included leaf, forage, pollen and silk and the mean levels of Cry3Bb1 ranged from 51 μg/g dry weight (MON 87410 pollen) to 400 μg/g dry weight (MON 87411 leaf). The mean CP4 EPSP S protein levels

wer e highest in root ( 81 and 66 μg/g dry weight for MON 87410 and MON 87411, respectively) and lowest in seed (means of 3.6 and 3.0 μg/g dry weight in MON 87410 and MON 87411, respectively)

. Similarly, the same tissues were evaluated for production of CP4 EP SPS protein and ranged from a mean of 14 μg/g dry weight (MON 87411 forage) to 43 μg/g dry weight in silk of both lines.

C. Toxicological Profile

Consistent with section 408(b) (2) (D) of the FFDCA, EPA has reviewed the available scientific data
and other relevant information in support of this action and considered its validity, completeness and reliability and the relationship of this information to human risk. EPA has also considered available information concerning the variability of the sensitivi ties of major identifiable subgroups of consumers, including infants and children.

D. Mammalian Toxicity and Allergenicity Assessment The Dv49 insecticidal nucleic acid component in MON 87410 and MON 87411 is currently being evaluated for safety and efficacy. Nucleic acids ( i.e ., RNA, dsRNA, and DNA) are present in all living organisms and are routinely consumed as a part of human and animal diets with no apparent adversePage 9 of 14

effects ( Parrott et al , 2010; Ivashuta et al ., 2009). The EPA recently affirmed (U.S. EPA, 2010b) that “DNA and RNA are common to all forms of plant and animal life, and the Agency knows of no instance
where these nucleic acids have been associated with toxic effects related to their consumption as a component of food”.

Double stran ded RNAs can be tailored to be highly specific to the target organism ( Whyard et al ., 2009 ) and the Dv49 dsRNA produced in MON 87410 and MON 87411was designed to achieve a high level of specificity ( i.e ., narrow spectrum of activity) resulting in a low lik elihood of off

- target effects and/or

non

- target activity. Due to these specificity characteristics, limited exposure under this EUP and
selected locations of the cultivated plots, adverse impacts on non

- target organisms are unlikely.

In addition, humans and other vertebrates lack the cellular “machinery” to appropriately process orally
consumed dsRNA to induce RNAi. So far, there are no documented cases of vertebrates being susceptible to ingested dsRNA. In addition, only a subset of invertebrates is susceptible to ingested
dsRNA. Thus, it is reasonable to assume that a single component of DV 49 will not cause adverse effects
in humans and animals.

Since the inserted Dv49 trait is expressed as a dsRNA and does not produce a novel pro tein, there is no need to consider potential allergenicity for Dv49. However, if one considers the criteria typically examined for allergenicity, the following information is available:

1. Source of the trait. Diabrotica virgifera ( Dv49 derived ) is not known to be a source of allergenic proteins. The source of the trait is composed of nuclei c acids ... specifically, double stranded ribonucleic acid (RNA). RNA is a ubiquitious family of large biological molecules that performs multiple vital roles in the c oding, decoding, regulation, and expression of genes. Together with DNA, RNA comprises the nucleic acids, which, along with proteins, constitutes
the three major macromolecules essential for all known forms of life.

2. Prevalence in food

. Nucleic acids ( i. e ., RNA, dsRNA, and DNA) are present in all living organisms and are routinely consumed as a part of human and animal diets with no apparent
adverse effects (Parrott et al , 2010; Ivashuta et al ., 2009). 0.1 % to 0.2% total RNA is siRNA soybean ( Ivashuta et al , 2009 ). Small dsRNA are abundant in soy, corn, and rice (Ivashuta et al

., 2009). In addition, many of these small RNAs are identical to vital mammalian genes.
3. Digestibility. Currently, there are no documented cases of vertebrates being susceptible t o ingested dsRNA. There is a lack of stability within the digestive tract and blood due in part to i) low pH depurinates nucleic acids; ii) RNA is degraded and quickly cleared from serum ( Behlke, MA, 2006) ; and iii) pancreatic and salivary nucleases metabolize dsRNA ( Park, NJ et al , 2006 ). Only a subset of inverte brates is susceptible to ingested dsRNA ( Terenius, O., et at , 2011 ).
4. Conclusion. Considering all of the available information, EPA has concluded that the potential
for Dv49 to be a food allergen is minimal

.

Aggregate Exposures

In examining aggregate exposure, EPA consider s available information concerning exposures from the pesticide residue in food and all other non

- occupational exposures, including drinking water from ground water or surface water and exposure through pesticide use in gardens, lawns, or buildings (reside ntial and other indoor uses).Page 10 of 14

The Agency has considered available information on the aggregate exposure levels of consumers (and major identifiable subgroups of consumers) to the pesticide chemical residue and to other related
substances. First, with r espect to general components of PIP, source of trait is composed of nucleic acid
which is defined under FIFRA as ribosides of

– adenine, guanine, cytosine, and uracil; and polymers of
ribose

- 5’

- monophosphates of uracil, cytosine, guainine, and adenine li nked by successive 3’

- 5’ phosphod iester bonds (also known as rib onucleic acid). In addition, an exemption from the requirement of tolerance has been established by the Agency with a history of safe consumption . § 174.507 Nucleic acid that are part of a PI P; exemption from the requirement of a tolerance. Residues of nucleic acids that are part of a plant

- incorporated protectant are exempt from the requirement of a tolerance [72 FR 20434, April 25, 2007]. Second, and specific to the RNA inhibitory (RNAi) m olecule comprising the Dv49 gene is double

- stranded ribonucleic acids (RNA) molecule. RNA is a component of normal diets with
an estimate of 0.1%

- 1% g RNA per day. In addition, small dsRNA are abundant in soy, corn, and rice (Ivashuta et al ., 2009). Exposure via the skin or inhalation is not likely since the plant

- incorporated protectant is contained within plant cells, which essentially eliminates these exposure routes or reduces these exposure routes to negligible

. Exposure via residential or lawn use to infants and children is also not expected because the use sites for the Dv49 dsRNA are all agricultural for control of insects. Oral exposure, at very low levels may occur from ingestion of processed corn products and, potentially, drinking water , i f corn stubble were not degraded by soil microbes

.

Cumulative Effects

Pursuant to FFDCA section 408(b)(2)(D)(v), EPA has considered available information on the
cumulative effects of such residues and other substances that have a common mechanism of toxicity.
These considerations included the cumulative effects on infants and children of such residues and other substances with a common mechanism of toxicity. Because there is no indication of mammalian toxicity to ingested nucleic acids, including tho se resulting from the plant

- incorporated protectant, we
conclude that there are no cumulative effects for the Dv49 dsRNA

. For information regarding EPA’s efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the
cumulati ve effects of such chemicals, see EPA’s website at http://www.epa.gov/pesticides/cumulative

.

Determination of Safety for U.S. Population, Infants and Children

Using biotechnology methods, Monsanto Company has deve loped MON 87410 and MON 87411 that confers resistance to corn rootworm (CRW; Diabrotica spp .). MON 87410 and MON 87411 contain a suppression cassette with an inverted repeat sequence (Dv49) derived from Diabrotica virgifera . The expression of the Dv49suppression cassette results in the formation of a double stranded RNA (dsRNA) transcript. Upon consumption by the CRW pest , the plant

- produced Dv49 dsRNA in MON 87410 and MON 87411 is recognized by the CRW’s RNA interference (RNAi) machinery resulting in the down

regulation of the targeted Snf7 gene expression and eventual CRW mortality.

The Dv49 insecticidal nucleic acid component in MON 87410 and MON 87411 is currently being evaluated for safety and efficacy. Nucleic acids (i.e., RNA, dsRNA , and DNA) are present in all living organisms and are routinely consumed as a part of human and animal diets with no apparent adverse effects (Parrott et al , 2010; Ivashuta et al

., 2009). Nucleic acids are considered to be “generally recognized as safe” ( GRAS) by the U.S. Food and Drug Administration (FDA) ( U.S. FDA 1992) and exempt from the requirement of a tolerance under the Federal Food Drug and Cosmetic Act by the U.S.

EPA . The EPA recently affirmed ( U.S. EPA, 2010b) that “DNA and RNA are common to all forms of plant and animal life, and the Agency knows of no instance where these nucleic acids have beenPage 11 of 14

associated with toxic effects related to their consumption as a component of food”. However, the mode
of action of DV49 against CRW will require a re

- examination of these assumptions on dietary safety before making a section 3 registration decision.

Finally, and specifically in regards to infants and children, FFDCA section 408(b)(2)(C) provides that
EPA shall asses s the available information about consumption patterns among infants and children, special susceptibility of infants and children to pesticide chemical residues, and the cumulative effects
on infants and children of the residues and other substances with a common mechanism of toxicity. In addition, FFDCA section 408(b)(2)(C) provides that EPA shall apply an additional tenfold margin of safety for infants and children in the case of threshold effects to account for prenatal and postnatal toxicity and the co mpleteness of the data base unless EPA determines that a different margin of safety
will be safe for infants and children.

Based on its review and consideration of all the available information, as discussed in more detail above, the Agency concludes that there is a reasonable certainty that no harm will result to the U.S. population, including infants and children, from aggre gate exposure to residues of the Dv49 dsRNA and the genetic material necessary for its production in corn. This includes all anticipated dietary exposures and all other exposures for which there is reliable information. The Agency has also concluded, agai n for the reasons discussed in more detail above, that there are no threshold effects of concern and, as a result, that an additional margin of safety for infants and children is unnecessary in this instance.

Other Considerations

Endocrine Disruptors As required by the Administrator under the Federal Food, Drug, and Cosmetic Act ( FFDCA) section 408(p), EPA has developed the Endocrine Disruptor Screening Program (EDSP) and has begun to implement the screening program that is to be used to test all pesti cides to determine whether certain substances (including pesticide active and other ingredients) may have an effect in humans or wildlife similar to an effect produced by a “naturally occurring estrogen, or other such endocrine effects as the

Administrator may designate.” FFDCA section 408(p)(4), authorizes the Administrator, by order, to exempt from the requirements of the Estrogenic Substances Screening Program a biologic substance or
other substance if a determination is made that the substance is antic ipated not to produce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. Between October 2009 and February 2010, EPA issued test orders/data call

- ins for the first group of 67 chemicals, which contain 58 pest icide active ingredients and nine inert ingredients. RNAi is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP.

The Agency believes that RNAi likely is a substance that would not produce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. As such, pursuant to Section 408(p)(4), EPA will determine in the future whether it can exempt RNAi from the req uirements of the Section 408(p) EDSP. In the event the Agency does determine to exempt this substance from the EDSP, an order will be issued. For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, future lists, the test guidelines and the Tier 1 screening battery, please visit our website: www.epa.gov/endo/

.

Supporting DataPage 12 of 14

The submitted studies to support the registrant’s EUP request for Dv49 dsRNA as expressed in Event MON87410 and MON 87411 are listed below with their respective MRID No. and classification (see Table 2). The Agency reviewed the submitted studies (including product characterization) and they are individually summarized in Data Evaluation Repor ts (DERs). Each DER contains the objective, test methods/materials, results and findings, conclusion, and classi fication of each study. This

me morandum represents the Agency’s overall recommendation for Monsanto EUP request based on the summary of data reviewed in each DER (for detailed information of each study, refer to the DER according to their respective assigned MRID number).

Table 2. Compilation of Submitted Data and Classification for MON 87410 and MON 87411 expressing Dv49 dsRNA protein [EPA. Reg. No. 524

- EUP

- RNU] in support of Monsanto’s EUP request

. MRID NO STUDY TITLE

48919001 Molecular characterization Data for MON 87410 and MON 87411 Supporting an

Experimental Use Permit to Test MON 87410, MON 87411, Combined Trait Insect Control Corn, Combinations Necessary for their Production , and Controls. ACCEPTABLE

48919002 Assessment of CP4 EPSPS and Cry3Bb1 Protein Levels in Corn Tissues from MON

87410 and MON 87411 Produced in a United States Greenhouse trial during 2011

. ACCEPTABLE

4891900 3 Assessment of DvS nf7 RNA levels in corn tissues from MON 87410 and MON 87411

Produced in a United States Greenhouse trial during 2011

. ACCEPTABLE

4891900 4 Preliminary Information to Characterize the Activity Spectrum of the dsRNA DvSnf7

Against a range of Insect Taxa. SUPPLEMENTAL

Table 3: Tolerance Exemptions
Protein/ Active Moiety Tolerance Exemption Information (a.i.)

40 CFR Date Crop(s)

Dv49 dsRNA §174.507 200 1/2007 All Corp PIPs Cry1A.105 §174.50 2 2008 Corn Cry2Ab2 §174.5 19 2008 Corn and Cotton Cry1F §174.5 20 200 1/2005 Corn and Cotton Vip3Aa20 §174.5 01/528 2008 Corn and Cotton Cry3Bb1 §174.5 18 200 4 Corn Cry34Ab1/ Cry35Ab1 §174.507 200 5 Corn

eCry3.1Ab §174.5 32 20 11

- 2013 Corn



### R EFERENCESPage 13 of 14

Baum, J.A., Bogaert, T., Clinton, W., Heck, G.R., Feldmann, P., et al. 2007. Control of coleopteran insect pests through RNA interference. Nat. Biotechnol 25: 1322

- 1326.

Behlke, M.A. (2006). Molecular Therapy 13: 644

- 670.

Betz, F.S., B.G. Hammond, and R.L. Fuchs. 2000. Safety and advantages of Bacillus thuringiensis protected plants to control insect pests. Regul. Toxicol. Pharmacol. 32:156

- 173.

Fire, A., Xu, SQ., Montgomery, M.K., Kostas, S.A., Driver, S.E, et al. 1998. Potent and specif ic genetic interference by double

- stranded RNA in Caenorhabditis elegans . Nature 391: 806

- 811.

Parrott, W., B. Chassy, J.

, Ligon, L. Meyer, J. Petrick, J. Zhou, R. Herman, B. Delaney and M. Levine. 2010. Application of food and feed safety assessment principles to evaluate transgenic approaches to gene modulation in crops. Food and Chemical Toxicology 48:1773

- 1790.

Park, N.J, Li, Y., Yu, T., Brinkman, B.M.N., and Wong, D.T. (2006). Clinical Chemistry 52: 988

- 994.

Ivashuta, S.I., J.S. Petrick, S.E. He isel, Y. Zhang, L. Guo, T.L. Reynolds, J.F. Rice, E. Allen and J.K. Roberts. 2009. Endogenous small RNAs in grain: Semi

- quantification and sequence homology to human and animal genes. Food and Chemical Toxicology 47:353

- 360.

Kim, D. W ., Sung, H., Shin, D. , Shen, H., Ahnn, J., et al . 2011. Differential physiological roles of ESCRT complexes in Caenorhabditis elegans

. Mol Cells 31: 585

- 592.

McClintock, J.T., C.R. Schaffer, and R.D. Sjoblad. 1995. A comparative review of the mammalian toxicity of Bacillus thuringiensis

- based pesticides. Pest. Sci. 45:95

- 105.

Sweeney, N.T., Brenman, J.E., Jan, Y.N., Gao, F.B. 2006. The coiled

- coil protein shrub controls neuronal morphogenesis in Drosophila . Curr Biol 16: 1006

- 1011.

Terenius, O., Papanicolaou, A., Garbutt, J.S., Eleftherianos, I., Huvenne, H., Kanginakudru, S., et al. (2011). Insect Physiology 57: 231

- 245.

U.S. EPA. 1998. Reregistration eligibility decision (RED) Bacillus thuringiensis. EPA738

- R

- 98

- 004. U.S. Environmental Protection Agency, Washington, D.C.

U.S. EPA. 2001. Bacillus thuringiensis Cry1F protein and the genetic material necessary for its production in corn; Exemption from the requirement of a tolerance. Federal Register 66:30321

- 30325.

U.S. EPA. 2005a. Bacillus thuringiensis Cry34Ab1 and Cry35Ab1 proteins and the genetic material necessary for their production in corn; Exemption from the requirement of a tolerance. Final Rule. Federal Register 70:55254

- 55260.

U.S. EPA. 2005b. Bacillus thuringiensis Cry34Ab1 and Cry35Ab1 proteins and the g enetic material necessary for their production (PHP17662 T

- DNA) in event DAS

- 59122

- 7 corn (OECD Unique
Identifier: DAS

- 59122

- 7). U.S. Environmental Protection Agency, Washington, D.C. http://www.epa.gov/pesticides/biopesticides/pips/cry3435ab1

- brad.pdfPage 14 of 14

[A ccessed March 29, 2012]

U.S. EPA, 2008a. Bacillus thuringiensis Cry1A.105 and Cry2Ab2 insecticidal proteins and the genetic material necessary for their production in corn [PC Codes 006515 (Cry2Ab2), 006514 (Cry1A.105)].
U.S. Environmental Protection Agency, Washington, D.C. http://www.epa.gov/pesticides/biopesticides/pips/mon

- 89034

- brad.pdf [Accessed March 29, 2012]

U.S. EPA, 2008b. Bacillus thuringiensis Cry1A.105 and Cry2Ab2 insecticidal proteins and the genetic material necessary for their product ion in corn [PC Codes 006515 (Cry2Ab2), 006514 (Cry1A.105)]. U.S. Environmental Protection Agency, Washington, D.C. http://www.epa.gov/pesticides/biopesticides/pips/mon

- 890 34

- brad.pdf [Accessed March 29, 2012]

U.S. EPA. 2009. Bacillus thuringiensis Vip3Aa20 insecticidal protein and the genetic material necessary for its production (via elements of vector pNOV1300) in event MIR162 maize (OECD Unique
Identifier: SYN

- IR162

- 4). U.S. Environmental Protection Agency, Washington, D.C. http://www.epa.gov/pesticides/biopesticides/ingredients/factsheets/factsheet_006599.html . [Accessed 3

- 29

- 2012]

U.S. EPA. 2010a. Cry1Ab and Cry1F Bacillus thuringiensis ( Bt ) corn plant

- incorporated protectants. U.S. Environmental Protection Agency, Washington, D.C. http://www.epa.gov/pesticides/biopesticides/pips/cry1f

- cry1ab

- brad.pdf [Accessed 3

- 29

- 2012].

U.S. EPA, 2010b. Bacillus thuringiensis Cry3Bb1 protein and the genetic material necessary for its production (vector PV

- ZMIR39) in MON 88017 corn (OECD Unique Identifier: MON

- 88Ø17

- 3). U.S. Environmental Protection Agency, Washington, D.C. http://www.epa.gov/pesticides/biopesticides/pips/cry3bb1

- brad.pdf [Accessed 3

- 29

- 2012]

U.S. FDA. 1992. Statement of policy: Foods derived from new plant varieties. Federal Register 57:
22984

- 23005.

Vaccari, T., Rusten, T.E., Menut, L., Nezis, I.P., Brech, A., et al. 2009. Comparative analysis of

ESCRT

- I, ESCRT

- II and ESCRT

- III function in Drosophila by efficient isolation of ESCRT mutants. J
Cell Sci 122: 2413

- 2423.

Whyard, S., A.D. Si ngh and S. Wong. 2009. Ingested double

- stranded RNAs can act as species

- specific insecticides. Insect Biochemistry and Molecular Biology 39:824

- 832.