CRISPR

What CRISPR is?

• CRISPR is a technology that can be used to edit genes and, as such,
will likely change the world.
• Before the CRISPR method was disclosed in 2012….. There were ways
to edit the genomes of some plants and animals
• Clustered regularly interspaced short palindromic repeats
Disadvantage
it took years and cost hundreds of thousands of dollars. CRISPR has
made it cheap and easy.
Advantages
• Transform medicine---- enabling us to not only treat but also prevent
many diseases.
• We may even decide to use it to change the genomes of our children.
• An attempt to do this in China has been condemned as premature
and unethical
Cas protein
• The main part of CRISPR is “Cas” proteins
• Found in bacteria where they help defend against viruses.
• The Cas9 protein is the most widely used by scientists.
• This protein can easily be programmed to find and bind to almost any
desired target sequence-----by giving it a piece of RNA to guide it in its
search.
How CRISPR works?
• 1. CRISPR Cas9 protein is added to a cell along with a piece of guide RNA
• 2. the Cas9 protein hooks up with the guide RNA and then moves along the
strands of DNA until it finds and binds to a 20-DNA-letter long sequence
that matches part of the guide RNA sequence.
• 3. The standard Cas9 protein cuts the DNA at the target.
• 4. When the cut is repaired, mutations are introduced that usually disable a
gene.
• It’s called genome editing – or gene editing – but usually the results are
not as precise as that term implies.
• CRISPR can also be used to make precise changes such as replacing faulty
genes – true genome editing – but this is far more difficult.
Advantages
• Modify genetic disease-causing genes in embryos-----in next
generations too
• Install genes that cause lifelong protection against infections
Disadvantages
• In 2016…James Clapper….termed genome editing as weapon of mass
destruction
• Unintentional misuse can cause economic and national security
concerns
• Genome altering unethical
• Can create biological weapons
Gene Therapy
• Gene therapy, the introduction of a normal gene into an individual’s genome in order to
repair a mutation that causes a genetic disease.
Approaches
1. Replacing defected gene with a healthy gene
2. Knocking out a mutated gene
3. Introducing new gene to treat the disease

When a normal gene is inserted into the nucleus of a mutant cell,

1. The gene most likely will integrate into a chromosomal site different from the
defective gene although that may repair the mutation
2. A new mutation may result if the normal gene integrates into another functional gene.
3. If the normal gene replaces the mutant allele, -----restored to the un diseased
phenotype.
Types
1. Somatic cells therapy ---Human gene therapy has been attempted
on somatic (body) cells for diseases such as cystic fibrosis, familial
hypercholesterolemia, cancer, and severe combined
immunodeficiency (SCID) syndrome-----the modification is not
passed on to the next generation.
2. Germline gene therapy corrects cells inside the germ line (e.g.,
cells of the ovary or testis)-----those cells will undergo meiosis and
provide a normal gametic contribution to the next generation.

Germline gene therapy has been achieved experimentally in animals

but not in humans.
Challenges
• Disease must be well-understood
• Gene identification
• A working copy of the gene must be available
• Efficient way to delivery healthy gene is available
• Genetic link of disease must be known
Procedure
• Carrier molecule required
• Vector is made from altered virus
• Vector should deliver desired gene to host cell without causing a
disease
• Once inserted the desired gene must restore the function of a protein
• Vector can be given intravenously or injected into specific tissue
Properties of vector
• Safe for use
• Should express in host ----as long as required
• Should not cause an allergic reaction
• Unrecognized by immune system
Techniques of delivering vector
• Ex-vivo
• In-vivo
Disadvantages
• Short-lived nature of gene therapy ----the rapid dividing nature of
cells halts it ----so patients have to undergo multiple gene therapies
• Immune response
• Problems with viral vectors….virus may recover its ability to cause
disease
• Insertional mutagenesis--- virus may target wrong cell---tumor
suppressor gene-could induce tumor
• Expensive