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DATA REVIEW RECORD:

Active Ingredient: dsRNA transcript comprising a Dv49 inverted repeat sequence derived 

from Diabrotica virgifera and the genetic material necessary for its 

production in corn (via plasmid vector PV-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 and MON 

87411 Produced in a United States Greenhouse trial during 2011. 

48919003 Assessment of DvSnf7 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.

BACKGROUND

Monsanto Company is seeking an Experimental Use Permit (EUP) to test a new plant-incorporated

protectant (PIP) technology, MON 87410 and MON 87411, combined 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 pests is an important component of producing highly 

efficacious products that also support durable Insect Resistance Management (IRM). The PIP events 

comprising the combined trait products 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. 67979-14; 

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-regulation of the targeted Snf7 (this gene is only in CRW) gene and 

eventual CRW mortality. MON 87410 and MON 87411 also produce the Cry3Bb1 protein to protect 
Page 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 Roundup1®

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-derived 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 evaluated by Monsanto for safety and efficacy. The PIP events comprising 

the single and combined trait products in this request include other 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 include Cry1A.105, Cry2Ab2, Cry 1F, 

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). Marker 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 experimental use permit (EUP) to conduct field testing and evaluate these 

single and combined events, corn PIP products that include two primary protocols spanning 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 includes 17,740 acres of PIP materials, 1,782 

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

for 2013-2014 is required 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 proposed planting includes field testing the two corn events (MON 87410 and MON 87411) 

representing a new class of PIPs active against CRW (Diabrotica spp.) with the 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], MON 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 information 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.
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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 and 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 event TC1507 is a truncated form of the native 

full-length Cry1F protein from Bacillus thuringiensis var. aizawai. The Cry1F protein does not

exhibit similarity 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 Maize 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 protects 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 Cry3Bb1 protein from this cassette was 

confirmed 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 in
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corn grain after heat treatment (U.S. EPA, 2008b). EPA reviewed safety assessment of the 

Cry2Ab2 protein as produced in MON 89034 and granted an 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 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, 2008a). EPA reviewed the 

safety assessment of the Cry1A.105 protein as produced in MON 89034 and granted 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 5307 

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 association 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 repeat sequence), Monsanto Company has submitted product characterization data which are 

reviewed in this report. 

RECOMMENDATION: 

 The 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 

Southern 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 sequences from PV-ZMIR10871 were detected in either 

MON 87410 or MON 87411.

48919001
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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 and MON 

87411 Produced in a 

United States Greenhouse 

Trial during 2011

The concentration of 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 weight 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 Coleoptera at exposure concentrations 

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

48919004
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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, Cry34Ab1/ 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 uses

as PIPs in major crops within U.S. agriculture. These PIPs have been combined through breeding for 

various assessments under this EUP. Two corn events within this proposed EUP are unregistered PIPs.

MON 87410 and MON 87411 produce the Cry3Bb1 protein present in the registered product 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 target 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). 

Product 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 expression 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 the 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 Dv49 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 

replication of the plasmid vector in bacteria and transfer of the T-DNA into plant cells. The inserted TDNA from this plasmid contains three gene cassettes.

1) The first cassette contain 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 as 
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the target mRNA to investigate the molecular 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 dsRNA into short interfering siRNAs 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 base pair (bp). Dv49 siRNA is 21-24 mer. High molecular weight (HMW) 

northern blot shows all detectable 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 cassette contains cry3Bb1 is a modified Bacillus thuringiensis (subsp. kumamotoensis) 

protein to protect against corn rootworm larval feeding.

3) The third cassette produces 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 Cry3Bb1 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 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 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 sensitivities 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 adverse 
Page 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 stranded 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 likelihood 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 DV49 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 protein, 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 nucleic acids... specifically, double 

stranded ribonucleic acid (RNA). RNA is a ubiquitious family of large biological molecules that 

performs multiple vital roles in the coding, 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 to 

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 invertebrates 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 considers 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 

(residential 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 respect 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 linked by successive 3’-5’ 

phosphodiester bonds (also known as ribonucleic 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 PIP; 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) molecule 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, if 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 those 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 

cumulative 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 developed 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 downregulation 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 been 
Page 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 assess 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 completeness 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 aggregate 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, again 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 pesticides 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 anticipated 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 pesticide 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 requirements 

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 Data
Page 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 Reports (DERs). Each DER contains the objective, test 

methods/materials, results and findings, conclusion, and classification of each study. This 

memorandum 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

48919003

Assessment of DvSnf7 RNA levels in corn tissues from MON 87410 and MON 87411 

Produced in a United States Greenhouse trial during 2011. ACCEPTABLE

48919004

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 2001/2007 All Corp PIPs

Cry1A.105 §174.502 2008 Corn

Cry2Ab2 §174.519 2008 Corn and Cotton

Cry1F §174.520 2001/2005 Corn and Cotton

Vip3Aa20 §174.501/528 2008 Corn and Cotton

Cry3Bb1 §174.518 2004 Corn

Cry34Ab1/

Cry35Ab1

§174.507 2005 Corn

eCry3.1Ab §174.532 2011-2013 Corn

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