TALENS

(Transcription activator-like effector

nukleases)
Kelompok 2

Fajri??

Ririn Sutanti (N012181018)

Transcription activator-like effector
nuclease (TALEN®) technology
leverages artificial restriction
enzymes generated by fusing a TAL
effector DNA-binding domain to a
DNA cleavage domain.
TALE DNA-binding domain
 TALE DNA-binding domain
• Tall effector are proteins secreted
by Xanthomonasbacteria via their type III
secretion system when they infect
various plantspecies.
• The DNA binding domain contains a repeated
highly conserved 33–34 amino acid sequence
with divergent 12th and 13th amino acids. These
two positions, referred to as the Repeat Variable
Diresidue (RVD), are highly variable and show a
strong correlation with specific nucleotide
recognition.
TALE DNA-binding domain
 DNA cleavage domain
• The non-specific DNA cleavage domain from
the end of the FokI endonuclease can be used
to construct hybrid nucleases that are active
in many different cell types
• The FokI domain functions as a dimer,
requiring two constructs with unique DNA
binding domains for sites in the target
genome with proper orientation and spacing.
TALEN mechanism
 TALEN mechanism
Workflow of genome editing of Your Favorite Gene
(YFG) using TALEN technology. The targent
sequence is identified, a corresponding TALEN
sequence is engineered and inserted into a
plasmid. The plasmid is inserted into the target
cell where it is translated to produce the
functional TALEN, which enters the nucleus and
binds and cleaves the target sequence.
Depending on the application, this can be used to
introduce an error (to knock out a target gene) or
to introduce a new DNA sequence into the target
gene.
 Application
• efficiently engineer stably modified human
embryonic stem cell and induced pluripotent
stem cell (IPSCs) clones and human erythroid
cell lines.
• it has been used in vitro to correct the genetic
defects that cause disorders such as sickle cell
disease, xeroderma pigmentosum, and
epidermolysis bullosa.