What are genome editing and CRISPR-Cas9?

: MedlinePlus Genetics
Last updated - 22/4/22
Possible Bias?

CRISPR-Cas9 was adapted from a naturally occurring genome editing system that
bacteria use as an immune defense. When infected with viruses, bacteria capture small
pieces of the viruses' DNA and insert them into their own DNA in a particular pattern to
create segments known as CRISPR arrays. The CRISPR arrays allow the bacteria to
"remember" the viruses (or closely related ones). If the viruses attack again, the bacteria
produce RNA segments from the CRISPR arrays that recognize and attach to specific
regions of the viruses' DNA. The bacteria then use Cas9 or a similar enzyme to cut the
DNA apart, which disables the virus.

Ethical concerns arise when genome editing, using technologies such as CRISPR-Cas9, is
used to alter human genomes. Most of the changes introduced with genome editing are
limited to somatic cells, which are cells other than egg and sperm cells (germline cells).
These changes are isolated to only certain tissues and are not passed from one
generation to the next. However, changes made to genes in egg or sperm cells or to the
genes of an embryo could be passed to future generations. Germline cell and embryo
genome editing bring up a number of ethical challenges, including whether it would be
permissible to use this technology to enhance normal human traits (such as height or
intelligence). Based on concerns about ethics and safety, germline cell and embryo
genome editing are currently illegal in the United States and many other countries.

How genome editing changed the world of large animal research

CRISPR/Cas9 might be the only genetic engineering technology where efficiency can be a
drawback, i.e., if only one allele of a gene should be inactivated and most samples
analyzed have a mutation on both alleles. Similarly, when editing is carried out in the
early embryo, mosaicism can occur. When working with mice, this is quickly bred out, but
for livestock, it could mean a 1- to 3-year delay, depending on the species. Using
homology-independent integration, to place transgenes or insert DNA fragments into the
genome, can result in the integration of concatemers. This may be advantageous if
additional copies improve transgene expression. If this is not the goal, then selection for
the correct single copy insertion is required.

Gene editing for pest control

Gene-editing has not yet been used as a conservation tool, but scientists are exploring
the possibility of using gene-editing tools to control invasive organisms. Current gene
editing options for pest control being explored are “sterile insects” and”gene drives”.

The Royal Society report focuses on how gene drives work. In normal sexual reproduction
50:50 chance governs which gene is inherited. In gene drives there is a much higher
chance that the edited gene is passed on. This increases the speed that the edited gene
spreads through a population.

A gene-drive, for pest control purposes, would target a gene “essential for viability or
fertility of the pest organism” and thereby reduce the population overtime.

There are two non-native wasp species in New Zealand, the common wasp and the
German wasp. Both have a significant and detrimental impact on our native flora and
fauna. Control methods for wasps are currently limited to small scale techniques. These
have make little impact given that 1 million hectares of beech forest are vulnerable to
wasp plagues.

One of the projects underway under the New Zealand’s Biological Heritage National
Science Challenge is investigating ways of “bringing Aotearoa’s invasive wasp problem
under control.” This project, includes sequencing the genomes of the two species,
alongside investigating other methods of biological control.

Scientist have not yet genetically modified wasps. However, they have used CRISPR with
honey bees successfully. It is likely that the technique will work with wasp species. A
further barrier is the complex social structure of wasp colonies. This complicates the
inheritance mechanism of a gene drive. Researchers will need to use computer modelling
to determine the potential impact of a gene drive. In addition, modelling will need to
evaluate how and where modified wasps could be released.

Wasps effectively hitchhiked their way to New Zealand. This means there is a possibility of
an altered wasp hitchhiking their way back to a region where wasps are an important
part of the ecosystem. The Royal Society states, “While New Zealand would greatly benefit
from eradication of these pests, their extinction here must not mean global extinction of
the entire species. “

The genetics and breeding of possums have been extensively studied in New Zealand
due to their impact on the indigenous ecosystems. Yet, there is still little known about
“functional genetics in marsupials” and a gene drive would be dependent on
considerable further research. As a first step, work is underway to sequence the genome
of possums.

If researchers successfully identified a gene for a gene drive, and then edited it
successfully, the application for conservation would have additional hurdles to overcome.
A huge breeding programme of altered possums would be required to introduce the
edited gene into the wild population at necessary densities. Initial calculations suggest a
quarter of a million altered possums released at a density of one per hectare.

Any research must also consider international impacts. While possums are an invasive
species in New Zealand, they are protected in Australia. If a gene altered possum was
introduced, intentionally or un-intentionally, to Australia it could be potentially
disastrous to Australian ecosystems.
Rats, unlike stoats and possums, are pests across the globe. This means that New
Zealand is not the only place with interest into the research into gene drives for rat
populations. Rats have been intensively studied as a lab species. The Norway rat was the
first mammal to have its genome completely sequenced. The ship rat has been less
studied. It has had its genome sequenced, the first step for developing a gene drive.

A gene drive in rats will be less challenging than other species, but still not easy. For
example, they are hard to genetically manipulate. Additionally, while scientists are making
progress, this will slow down the advancement of a gene drive.

Globally, a further issue is the potential for altered rats to hybridise with other related
species. In overseas contexts, rodents can be crucial parts of the ecosystem for example
as pollinators.

Scientists know little about the genetics of stoats. Developing a gene drive for stoats
could springboard the research done into the genetics and breeding of a closely related
species, mink.

A successful gene drive, like with possums and rats, would require a significant number of
altered species being released into the wild.

GENE EDITING SCENARIOS IN PEST CONTROL

August 2019

A technique called CRISPR has increased the speed, ease and accuracy of gene editing.
Modified from a system found in bacteria to cut up invading virus DNA, CRISPR is much
more precise than earlier gene editing techniques. However, this ability to edit genes is, in
many cases, ahead of our understanding of everything that different genes do, resulting
in the possibility of unintended effects.

As for wasps, genetically modified animals are defined as new organisms under the HSNO
Act, and therefore possums containing gene drive would be classified as ‘new organisms’.
As with wasps, risk assessments of organisms produced through gene drives are carried
out under the provisions of the HSNO Act on a case-by-case basis by the Environmental
Protection Authority.

Inputs and outputs of gene drive techniques will be regulated by the Hazardous
Substances and New Organisms Act (HSNO Act) if they come within the definition of an
‘organism’ and ‘new organism’ in this Act. ‘Organism’ is widely defined in the Act and
includes a genetic structure (other than a human cell) that is capable of replicating itself.
The definition of ‘new organism’ includes organisms belonging to species that were not
present in New Zealand prior to July 1998 and Genetically Modified Organisms (GMOs).
The definition of a GMO is very broad (organisms whose genes or genetic material have
been modified by in vitro techniques). Genetically modified animals are defined as new
organisms under the HSNO Act, and therefore gene edited wasps would be classified as
‘new organisms’

Gene drives offer a control method that is less harmful at an individual level than
conventional pest control methods, which usually involve killing the animal. However, this
is dependent upon being as harmless as possible in its effects on any ecological changes
the gene drive bring

Genetic pest controls and how they sit with Māori - Biological Heritage NZ

Published - 21/12/21

Originally from Eurasia, the German wasp and common wasp are becoming increasingly
common throughout Aotearoa, reaching extremely high densities of up to 40 nests per
hectare. A thriving nest can contain more than 20,000 wasps, meaning around 800,000
wasps could be living within the space of a rugby field.
That’s a lot of wasps.
They threaten taonga species by preying on native invertebrates and leaving less food
available for our birds. Because of this, high densities can have huge knock-on effects for
the rest of the ecosystem.
Symon says in te ao Māori (the Māori worldview), ngārara (insects and reptiles) are
culturally symbolic, embodying whakapapa that connects to Atua (deities) like Tāne-
mahuta (deity of forests and birds).

Wasps also threaten human health and are estimated to cost New Zealand about $133
million annually.
Symon says “There is a clear need to develop highly targeted, environmentally safe, and
socially and culturally appropriate approaches for the control of wasps. This could then
provide a roadmap for other pests.”

Gene drive and RNAi technologies: a bio-cultural review of next-generation tools

Published - 14/10/21

Comprehensive review of CRISPR-based gene editing: mechanisms, challenges,

Published - 9/1/24
Edited - 27/2/24