Teacher Notes for "Genetic Engineering Challenge

How can scientists develop a type of rice that could prevent vitamin A deficiency?"1
This analysis and discussion activity begins with an introduction to vitamin A deficiency, rice
seeds, and genetic engineering. Next, several questions challenge students to design a basic plan
that could produce a genetically engineered rice plant that makes rice grains that contain provitamin A. Subsequent information and questions guide students in developing an understanding
of the basic techniques of genetic engineering. Students use fundamental molecular biology
concepts as they think about how to solve a practical problem. This activity can be used to
introduce students to genetic engineering or to reinforce basic understanding of genetic
engineering.
Before students begin this activity, they should have a basic understanding of DNA, proteins, and
transcription and translation. To provide this background, you may want to use "From Gene to
Protein Transcription and Translation" (http://serendip.brynmawr.edu/sci_edu/waldron/#trans).
These Teacher Notes propose an extension activity to introduce students to some additional
complexities of the molecular procedures used in genetic engineering and a follow-up activity to
engage students in evaluating the arguments and evidence in favor of and opposed to the
development of genetically engineered Golden Rice.
Learning Goals
Promote student understanding of molecular biology concepts, e.g.:
Genes code for proteins (including enzymes).
The genetic code is universal.
Transcription of genes is the first step in producing proteins.
(Almost) all the cells in an organism have the same genes in their DNA, but different
types of cells have different amounts of the various proteins, which results in the different
characteristics of the different types of cells.
Differences in the rate of transcription of specific genes are a major cause of the differing
amounts of specific proteins in different types of cells.
Promoters at the beginning of each gene play a crucial role in regulating the rate of
transcription of each gene in different types of cells.
Introduce genetic engineering concepts, including plasmids and recombinant DNA
Develop an understanding of the basic steps of genetic engineering to produce genetically
modified food plants
In accord with the Next Generation Science Standards2:
Students prepare for the Performance Expectation HS-LS3-1. "Ask questions to clarify
relationships about the role of DNA and chromosomes encoding the instructions for
characteristic traits passed from parents to offspring."
Students learn the following Disciplinary Core Ideas:
"Genes are regions in the DNA that contain the instructions that code for the formation of
proteins." (LS1.A Structure and Function)
1 By Dr. Ingrid Waldron, Department of Biology, University of Pennsylvania, 2014. These Teacher Notes and the related Student Handout
are available at http://serendip.brynmawr.edu/exchange/bioactivities/geneticengineer.

2 Next Generation Science Standards, available at http://www.nextgenscience.org/next-generation-science-standards

1

"The instructions for forming species' characteristics are carried in DNA. Not all DNA
codes for a protein; some segments of DNA are involved in regulatory or structural
functions " (LS3.A Inheritance of Traits)
Students engage in recommended Scientific Practices, including "constructing explanations
(for science) and designing solutions (for engineering)".
This activity provides the opportunity to discuss two Crosscutting Concepts: "Cause and
effect: Mechanism and explanation" and "Structure and function".

This activity will also help students meet Common Core English Language Arts Standards for
Science and Technical Subjects, including "determine the central ideas or conclusions of the text;
summarize complex concepts, processes or information presented in a text by paraphrasing them
in simpler but still accurate terms".3
Suggestions for Discussion and Background Information
To maximize student participation and learning, I suggest that you have your students work
individually or in pairs to complete groups of related questions and then have a class discussion
after each group of related questions. In each discussion, you can probe student thinking and help
them develop a sound understanding of the concepts and information covered before moving on
to the next group of related questions. I recommend that you have a class discussion after
question 4, at the end of the section on "Inserting the Desired Genes in the DNA of Rice Plants",
and at the end of the section on "Ensuring that the Genes for the Enzymes to Make Pro-Vitamin
A are Active in Rice Grain Cells".
A key for this activity is available upon request to Ingrid Waldron, iwaldron@sas.upenn.edu.
Some additional background information and suggestions for discussion are provided below.
Pro-vitamin A, also called beta-carotene, is found in many plant foods (e.g. deep orange and dark
green vegetables such as carrots, sweet potatoes and spinach). As shown in the following
diagram, beta-carotene can be enzymatically split to form retinal (a form of vitamin A that can be
converted to the other forms of vitamin A used in the body). Vitamin A is found in some animal
foods (especially liver). Excess vitamin A in the diet can be toxic. Pro-vitamin A may be a safer
dietary source of vitamin A since increased intake of beta-carotene results in decreased
conversion of beta-carotene to vitamin A.

Vitamin A deficiency is widespread in children in poor countries, resulting in an estimated

250,000-500,000 cases of blindness and up to 2.5 million deaths each year.

3 From Common Core Standards Initiative, http://www.corestandards.org/ELA-Literacy/RST/11-12

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White rice has been milled and polished to remove the hull, bran and germ, leaving only the
endosperm which contains thousands of cells, abundant starch and some protein. In the figure in
the Student Handout, the endosperm is labeled as "white rice". The major advantage of white rice
is the removal of most of the oils, which tend to become rancid when stored, especially at warm
temperatures. Brown rice has the hull removed, but keeps the germ and bran. The major
advantage of brown rice is higher levels of B vitamins and vitamin E. Neither white rice nor
brown rice provides pro-vitamin A or vitamin A.
Question 2 will serve to remind the students that genes code for proteins, including enzymes that
synthesize other needed molecules such as pro-vitamin A.
With respect to question 3, genetic engineering to produce recombinant DNA in transgenic
organisms is only useful because the genetic code is universal, so any type of organism can
transcribe and translate a gene from any other type of organism.
Question 4 is designed to get students thinking about the problems that scientists confront when
developing a genetically engineered plant. Question 4a may be challenging for students if they
do not have any background information about genetic engineering; it is hoped that having
students discuss this question in pairs will stimulate them to come up with some possible
strategy, even if it is only injecting DNA in the rice cells' nuclei. With respect to question 4b,
some students may recognize that it will be easier to transform a small group of embryonic cells
that can replicate the inserted gene every time the cells divide and thus produce a rice plant that
will have the inserted gene in every one of the thousands of cells in each rice grain.
(continued)

The bacterium discussed in the section "Inserting the Desired Genes in the DNA of Rice Plants"
is Agrobacterium tumefaciens. This bacterium can inject a crucial part of the DNA in its plasmid
into plant cells to form recombinant DNA in the plant cell nucleus. The bacterial part of this
transgenic DNA contains genes that code for enzymes to make opines (modified amino acids that
are synthesized by the plant cells and leak out where they are consumed by the bacteria as food)
and genes that code for the production of plant hormones that stimulate plant cell division and
the production of Crown galls (as shown below; the plasmid is called Ti or tumor-inducing
because it induces the formation of Crown galls). These Crown galls provide abundant food for
the bacteria to grow and multiply. To make use of the genetic engineering capabilities of this
bacterium, scientists need to insert the genes for the enzymes to make pro-vitamin A into the TDNA of the Ti plasmid and remove the Crown gall-inducing bacterial genes from the T-DNA of
the Ti plasmid.

http://www.btny.purdue.edu/Extension/Pathology/PHM/BD/picts/agrobacteriumlc.jpg

Useful sources of additional information are

"Gene manipulation in plants" (http://www.open.edu/openlearn/science-mathstechnology/science/biology/gene-manipulation-plants/content-section-2.1)
"The Microbial World: Biology and Control of Crown Gall"
(http://archive.bio.ed.ac.uk/jdeacon/microbes/crown.htm).
This is just one example of how scientists frequently make use of the specialized capabilities of
biological systems to carry out genetic engineering. Another example is the use of viruses to
carry DNA into human cells for gene therapy.

The section, Ensuring that the Genes for the Enzymes to Make Pro-Vitamin A are Active in
Rice Grain Cells, engages students in thinking about the regulation of genes; this introduces an
important general concept and also provides the basis for understanding one aspect of genetic
engineering. The specific promoter used for genetically engineering rice plants that produce provitamin A in rice grains is a promoter for an endosperm-specific storage protein; this ensures that
the transformed genes are transcribed in rice grains.
Depending on your students, you may want to use the following diagram to discuss the role of
the promoter. Of course, the regulation of genes in eukaryotic cells involves much more than just
a promoter, but it is the promoter that needs to be incorporated at the beginning of the coding
region of the gene as it is prepared for use in genetic engineering.4

http://www.citruscollege.edu/lc/archive/biology/PublishingImages/0262l.gif

The class discussion of student responses to question 12 should link back to issues, concepts and
problems that were included in your discussion of question 4. You may find the figure on the
next page useful for this discussion, although it shows some additional information not included
in the Student Handout and the plant it shows is not a rice plant. If you have your students
complete the Extension Activity (see next page), your class discussion of this extension activity
should refer back to your discussion of question 12. Even if you do not have your students
complete the Extension Activity, you may want to read the second reference, which provides
helpful additional information.
4 Additional information about preparing the genes for genetic engineering is provided in "How to Make Transgenic Plants:
Animation Demo" (http://cls.casa.colostate.edu/transgeniccrops/animation.html). Explanations of the regulation of transcription
are available at http://www.nature.com/scitable/topicpage/gene-expression-14121669 and
http://sandwalk.blogspot.com/2008/09/how-rna-polymerase-binds-to-dna.html An excellent brief video description of regulation
of eukaryotic DNA transcription is available at http://www.hhmi.org/biointeractive/regulation-eukaryotic-dna-transcription.
"Regulating Genes" (http://www.pbs.org/wgbh/nova/education/interactives/regulating-genes/) is a useful introduction to Evo
Devo (how changes in gene regulation influence development and how these changes have contributed to evolution).

(http://classes.midlandstech.edu/carterp/Courses/bio225/chap09/09-18_TiPlasmid_1.jpg )

Extension Activity
This extension activity is shown on the last page of these Teacher Notes. This extension activity
introduces students to many of the complexities and specific molecular procedures used in
genetic engineering, including:
the use of restriction enzymes to make recombinant plasmids and the use of transgenic
bacteria to clone the gene of interest
the need for a marker that will allow scientists to select for plant cells that have been
transformed
the need for a termination sequence for the coding region of the gene
an alternative method of transforming plant cells (biolistic transformation using a gene
gun)
the need to cross the transgenic rice plants with local breeds that have desirable
characteristics such as adaptation to local weather conditions and resistance to a variety
of plant pests and diseases.
Some additional steps that are needed before a genetically engineered food plant can be released
for agricultural use (e.g. testing the safety of the food and testing for possible adverse
environmental effects) are included in Extension Activity 2.
The second recommended source in Extension Activity 1 also illustrates the iterative nature of
scientific research; this reading describes a few of the multiple steps involved in continuously
improving the effectiveness of the genetic engineering techniques by repeatedly trying different
approaches and using the results of these tests, together with new ideas, to develop new
approaches to be tested. This iterative process has included:
trying different techniques to transform rice plant cells,
research on the biosynthetic pathway for producing beta-carotene which has shown that
only two enzymes need to be genetically engineered into the rice plant,
6

research to find specific versions of the genes for these enzymes that result in production
of higher levels of beta-carotene.
Using this extension activity will also illustrate the iterative nature of learning; we need to begin
with some basics, including a basic conceptual framework, and then repeatedly improve our
understanding by incorporating new information and concepts with our previous understanding.
Additional information about some of the technical aspects of genetic engineering that are not
included in this activity is available from:
"Development of Recombinant DNA" (http://ocw.mit.edu/courses/biology/7-01scfundamentals-of-biology-fall-2011/recombinant-dna/development-of-recombinant-dna/)
"Basic Mechanics of Cloning: Restriction Enzymes and Cloning Vectors"
(http://ocw.mit.edu/courses/biology/7-01sc-fundamentals-of-biology-fall-2011/recombinantdna/basic-mechanics-of-cloning/)
Follow-up Activity
The genetically engineered rice plants that produce pro-vitamin A in rice grains are called
Golden Rice. The discussion activity, "Golden Rice Evaluating the Pros and Cons" (available
at http://serendip.brynmawr.edu/exchange/bioactivities/GoldenRice), engages students in
evaluating the evidence and arguments related to Golden Rice and other possible strategies for
preventing vitamin A deficiency. Students use this information to develop evidence-based
conclusions about Golden Rice and other proposed strategies. Students also develop questions
that could provide important additional information for evaluating the arguments in favor of and
opposed to Golden Rice and other policy proposals. In addition, students analyze how two
reasonably accurate articles can present totally opposing points of view on a complex policy
issue.
Resources for Teaching about Other Types of Genetic Engineering
Cloning, including "What is cloning?", "Click and Clone", "Why clone?", "Cloning Myths",
"What are the risks of cloning?", etc. (http://learn.genetics.utah.edu/content/tech/cloning/)
Gene Therapy: Molecular Bandage?, including "What is gene therapy?", "Gene Delivery:
The Key to Gene Therapy", "Cystic Fibrosis: Case Study", "Tools of the Trade", Challenges
in Gene Therapy", etc. (http://learn.genetics.utah.edu/content/tech/genetherapy/)

Extension Activity 1 Learning More about How Scientists Do Genetic

Engineering
Scientists have developed a genetically engineered type of rice called
Golden Rice which produces rice grains with substantial quantities of provitamin A. The actual process has been significantly more complex than the
basic steps described in the activity you have completed. For useful
additional information read and view:
"How do you make a transgenic plant?" (including an animation demo
from the University of Nebraska at Lincoln;
http://cls.casa.colostate.edu/transgeniccrops/how.html)
(http://cls.casa.colostate.edu/transgeniccrops/animation.html)
"Gene Manipulation in Plants" (http://www.open.edu/openlearn/sciencemaths-technology/science/biology/gene-manipulation-plants/contentsection-4.3)
As you read these sources, note any additional steps or modifications of the
procedures needed to genetically engineer rice plants that produce provitamin A in their rice grains.
Prepare a revised and expanded description of the steps needed to
produce rice plants that make significant quantities of pro-vitamin A in
their rice grains.