“Fixing” Gene Expression:

Central Dogma and Genetic Medicine

(from HHMI BioInteractive.org)

Part 1: Card Sorting

“Fixing” Gene Expression
The flow of information from DNA to RNA to protein represents
the way most genes are expressed in eukaryotic cells. It is also
referred to as central dogma of molecular biology. You will
review the central dogma by sorting cards that illustrate the
molecules involved in transcription, RNA processing, and
translation. You will then suggest ways to intervene in this
pathway to treat different genetic conditions, including
hemophilia, cystic fibrosis, Huntington’s disease, and sickle cell
disease. The cards are at the end of this handout. You can either
print them and sort them into the correct order, or just look at
them and determine their order.

List here the correct order of the cards: Click or tap here to enter text.

Part 1: Click & Learn

Go to: https://media.hhmi.org/biointeractive/click/genetic-medicine-interactive/

As you proceed through the Click & Learn “Central Dogma and Genetic Medicine,” follow the
instructions and answer the following questions.

1. Let’s review! The central dogma of molecular biology refers to the process of gene expression.
Write the definition of gene expression in your own words.

Gene expression is the process of converting genetic material into proteins.

2. Click on the “Central Dogma” menu tab at the top of the screen. For each step, click on the tab
or scroll through the page as it outlines the steps in eukaryotic gene expression and briefly
summarize each step with the following information. The steps are: Transcription, RNA Splicing,
mRNA Transport, Translation, and Protein Processing.

Gene Expression Steps Molecules Involved (What molecules Summary (What happens
and proteins are involved in this step? during this step?
Transcription RNA polymerase and DNA RNA polymerase
transcribes DNA into a
 functional DNA transcript
that is complementary to
the DNA sequence.
RNA Splicing Spliceosomes and DNA transcript Spliceosomes remove
introns and connect exons
in the DNA transcript to
create mRNA.
mRNA Transport mRNA mRNA is moved from the
nucleus to the cytoplasm
of the cell.
Translation Ribosome, mRNA, tRNA and amino Ribosomes translate the
acids mRNA using tRNA to
create amino acids that
form a polypeptide chain.
Protein Processing Polypeptide chain The polypeptide chain is
folded into a real 3-
deminsional object of a
protein.

3. Mutations in the DNA can affect the structure and functions of proteins. Some mutations may
even cause genetic diseases. Scientists and doctors can intervene at different points during gene
expression to develop treatments for such genetic diseases – or genetic medicine. Let’s learn
about the genetic medicines that are being developed.

Select the “Genetic Medicine” tab located on the top right of the screen. Click the tab
corresponding to the genetic medicine(s). Scroll through the interactive and click on the pink “+”
sign labeled with each genetic medicine. Read the “Genetic Medicine” tab material, watch the
video, and read the information in the “Learn More” link. Then, briefly summarize each one with
the following information.

Genetic Medicine Short Summary (Write a Detailed Description (Describe how this
once-sentence summary genetic medicine would be used to treat a
of how this genetic genetic disease (For example, mention how
medicine works.) it would fix the disease-causing mutation
and/or result in a functioning protein.)
CRISPR-Cas9 CRISPR-Cas9 is used to CRISPR-Cas9 has the possibility of fixing bad
be able to change DNA mutations to treat disease.
in a specific location.
Gene Therapy A viral vector is The virus can alter protein function by
commonly used to replacing harmful places of the genome
deliver functioning with functioning genes to treat disease.
versions of genes to the
cell.
Gene Switches These are non-coding Altering or activating/deactivating these
parts of the genome gene switches can alter gene expression in
that regulate gene diseases.
expression.
 Exon Skipping RNA binds to a RNA
transcript to alter the
splicing of the
transcript.
RNA Interference RISC prevents the
expression of a specific
region of mRNA by
binding to it.
Small Molecule Drug Small molecule drugs
are easily absorbed into
cells and taken up due
to their size.
Altering slicing allows for exons to be
removed or skipped over to alter possible
disease causing regions.
Preventing the expression of genes can
cause the suppression of certain disease
causing proteins to be created.
This allows for small molecules to interact
directly and quickly by altering disease
linked regions of the genome.

4. Now let’s learn about some of the disease that may be treated using these genetic medicines.
Scroll through the interactive and click on the pink “+” sign that is labeled with the diseases.
Chose two of the five diseases for your group to work on. Next, click on the “Case Study” tab at
the top to reveal information about the disease. Reach the “Case Study” material, watch the
video, and read the information in the “Learn more” link. Then provide the following for each of
the five diseases: Leber Congenital Amaurosis, Sickle Cell Disease, Duchenne Muscular
Dystrophy, Huntington’s Disease, and Cystic Fibrosis
a. What are the key characteristics of the disease and whom does it affect?

Leber Congenital Amaurosis causes extreme far-sightedness or blindness at birth in infants. It

affects the retina’s signal to the brain when taking in light. Sickle Cell disease is a disease
resulting from a mutation on the gene for HBB. Those with the disease produce sickle shaped
red blood cells. Duchenne Muscular Dystrophy is a disease caused by mutations on the genes
that code for muscle protein dystrophin in muscle cells. This causes muscles to fail to adhere to
surrounding tissue which leads to wheelchair use and later premature death. Huntington’s
disease is caused by a mutation in the huntington gene with codes for normal nerve function.
This causes brain cell death overtime in people in adulthood through old age which causes
dementia and eventual death. Cystic Fibrosis is caused by one of the many genes that code for
the CFTR which causes an issue with Chloride ion diffusion and transport which usually leads to
infections.

b. How can the featured genetic medicine be used to treat the disease?

Gene therapy has been successfully used to treat Leber Congenital Amaurosis by targeting one
of the many possible causal genes RPE65. The gene switch for producing normal fetal
hemoglobin has been looked into for those with Sickle Cell Disease in order to continue
production of healthy red blood cells. The exon-skipping drug eteplirsen can cause the exon
containing the disease-causing mutation to be spliced out of the dystrophin mRNA. This allows
for a more functional protein. There is RNA interface technology being currently tested to treat
Huntington’s disease by reducing the production of the mutant huntington gene. There are
currently two small molecule drugs available to treat Cystic Fibrosis by targeting the genes
associated.
Questions 5-7 present disease scenarios. For each one you are asked to identify a way to treat the
disease by targeting a specific molecule or step in the central dogma pathway. Refer to your cards as
you think of an answer and make sure you pick a different approach for each disease.

5. Cystic fibrosis is a devastating illness that affects the lungs, pancreas, and intestines. In 1989,
researchers discovered that the disease is caused by a mutation in a gene that produces a
protein that channels chloride across cellular membranes. People with two copies (or alleles) of
the mutated gene have a buildup of mucus in the lungs, intestines, and other organs due to
nonfunctioning or absent channel proteins. Suggest two ways you could intervene to treat the
disease by targeting the DNA molecule and justify why each approach could be effective.

A possible treatment could be using CRISPR-Cas9 technology to edit the DNA sequence by getting rid
of one or both of the mutated alleles. Another possible treatment could be using gene therapy by
introducing a normal cell’s genes into the DNA.

6. Like cystic fibrosis, sickle cell anemia is an autosomal recessive condition. It can be caused by
mutations in the gene for β-globin (HBB). HBB is one of the two subunits of adult hemoglobin,
the protein that carries oxygen in red blood cells. People who inherit two copies of the mutation
produce abnormal hemoglobin, and their tissues are starved of oxygen. One interesting finding
is that some individuals with HBB mutations do not have sickle cell anemia because they have
another mutation that allows them to produce fetal hemoglobin throughout their lives. Fetal
hemoglobin production is normally turned off after birth. Based on this knowledge, suggest two
ways you could treat sickle cell anemia by targeting the transcription step of the fetal
hemoglobin gene and justify why each approach might be effective.

Introducing the needed activators to begin transcription would be a method to countering this.
Another option would be to introduce something that would ensure that anything that would
repress transcription would be removed.

7. Another disease caused by a mutation in a single gene is Huntington’s disease (HD), an

autosomal dominant condition. It is caused by mutations in a gene required for normal nerve
cell function. The mutations cause abnormal proteins to be produced and “stick” together and
accumulate in nerve cells, eventually interfering with normal cell operations. Suggest two ways
you could treat the disease by targeting the translation step for the HD protein and justify why
each approach might be effective.

Blocking the mRNA from being made would prevent this. Destroying the mRNA that itself would also
prevent this.

Part 3: Apply what you have learned

Hemophilia
Hemophilia is a rare genetic disorder in which the blood does not clot properly because it lacks sufficient
blood-clotting proteins. The disease is caused by mutations in a gene on the X chromosome. Because
males typically have only one X chromosome, to inherit the disease, they only need to inherit one copy
of the mutated gene, whereas females with two copies of the X chromosome must inherit two copies of
the mutated gene. Symptoms of the disease vary but generally include excessive internal and external
bleeding. Currently, there is no long-term cure, and many patients receive infusions of blood-clotting
proteins. Because the disease is caused by mutations in the DNA, scientists think that they could
someday treat it by intervening in the steps that occur from DNA to protein production.

Identify two ways a researcher could design an intervention to treat hemophilia. Provide a brief
explanation of each and justify why each approach might be effective.

Using gene therapy to introduce a normal healthy gene into blood cells containing clotting factors would
assist in reducing mutated cells. Using CRISPR-Cas9 to edit the gene to match a normal healthy blood
cell containing clotting factors could also be a helpful treatment.

Hutchinson-Gilford Progeria Syndrome

You are a researcher working on a treatment for Hutchinson-Gilford progeria syndrome, and extremely
rare genetic disorder that causes accelerated aging in children. Children with progeria generally appear
healthy at birth but soon start growing more slowly than other children and lose their hair. Additional
symptoms include stiffness of joints, heart problems, and stroke. These children typically die of heart
disease at an average age of 13 years.

Progeria is caused by a mutation in a single gene, called lamin A. Scientists have identified over 1,400
mutations in the lamin A gene that result in changes in transcription, RNA splicing, and/or protein
production. Lamin A codes for a protein required for the structural support of the nuclear envelop in
cells. Without a functional protein, the nuclear membrane becomes unstable, eventually damaging the
nucleus and causing cells to die.

Based on what you learned in the Click & Learn, propose a genetic medicine strategy you could develop
to treat patients with progeria. Describe which step in gene expression you might target and why you
would target that step, the intervention tool you would use, and explain how this strategy would treat
the disease.

Click or tap here to enter text.