BASE Editing

Base editing is a genome reediting approach that uses components

from CRISPR systems together with other enzymes to directly install
point mutations into cellular DNA or RNA without making double-
stranded DNA breaks (DSBs).
CRISPR systems: A single RNA, consisting of a crRNA sequence that is specific to the DNA
target. A tracrRNA sequence that interacts with the Cas9 protein, binds to a recombinant
form of Cas9 protein that has DNA endonuclease activity. The most amazing feature of
CRISPR is that the scissors can be programmed to search for, bind to and cut only a specific
DNA sequence.

The science behind the technology. All current base editors are comprised of the
HOW DOES BASE EDITING WORK?
cytidine deaminase enzyme and are therefore
only capable of C to T or G to A DNA base
transformations.

Since cytidine deaminase enzyme can only bind

A schematic showing R-loop formation by the base editors
and the interaction between the cytidine deaminase enzyme
and ssDNA
to single-stranded DNA, the local denaturation
of the target DNA upon dCas9:sgRNA binding (R-
loop formation). As in CRISPR, the specific locus
targeted with sgRNA must be followed by a
protospacer adjacent motif to allow for efficient
Cas9 binding.
In the structure of the Cas9 R-loop complex, the
eleven nucleotides furthest from the PAM on the
nontarget strand are disordered, suggesting they
are unencumbered and accessible for base
editing.

Once the R-loop forms, the cytidine deaminase

Depiction of the initial, intermediate, and final DNA
sequences involved in base editing, as well as a schematic of
the chemical reaction being catalyzed by the deaminase
enzyme.
enzyme directly binds the target nucleotide (C)
and chemically converts it to U. The resulting
U:G mismatch is then processed by cellular DNA
replication or repair, and resolves into a T:A
base pair. The overall DNA transformation is,
therefore, C:G to T:A.

The advantage of the technology.

WHAT IS A TECHNOLOGY DESIGNED TO DO?

Base editors have also been used in animals to reverse the consequence of tyrosinemia,
beta-thalassemia, muscular dystrophy, phenylketonuria, congenital deafness and a
type of cardiovascular disease in each case, by directly correcting a point mutation that
causes or contributes to the disease. In plants, base editors have been used to
introduce individual single DNA letter changes that could lead to better crops.

The disadvantages or ethical considerations

with the use of this technology.
The base editing may have the possibility of off-target effects of edits in the wrong
place and mosaicism when some cells carry the edit but others do not.
The people maybe will start using it for non-therapeutic and enhancement purposes, it
will harm the world ecology and increase the crime rate.
The parents will have the chance to make many decisions that affect their future
children, by edit the genes of the embryo and future generations.
YOLANDA LIN