TALENs animal models

NegotiableUpdate on 03/04

Model

Nature of the Manufacturer: Producers

Product Category

Place of Origin

Online Consult

Overview

TALENs animal model construction is an experimental animal model constructed using TALENs gene editing technology. TALENs can specifically recognize and cleave target DNA sequences by fusing the DNA binding domain of transcription activator like effectors (TALE) with the cleavage domain of nucleases (FokI), thereby achieving precise genome editing.

Product Details

TALENs animal models

1、 TALENs Technology Principles and Core Breakthroughs

（1） Structural design and mechanism of action

TALENs (Transcription Activation Like Effects or Nucleies) are a type ofArtificially engineered nucleasesIt consists of two parts:

DNA recognition domain (TALE protein)：

fromA repeating unit of 34 amino acidsComposition, each unit passes throughVariable amino acids (RVDs) at positions 12 and 13Specific recognition of a single base:

RVD type	Identify bases	binding strength
NI	A	high specificity
NG	T	high specificity
HD	C	high specificity
NN	G/A	Cross recognition

The length of the recognition sequence is usually15-20 bpEnsure targeting specificity.

DNA cleavage domain (FokI nuclease)：

Need to formdimerTo cut DNA, TALENs need to be designed in pairs (left and right arms), with the cutting site located between the two arms (usually the spacer is 12-20 bp).

（2） Gene editing mechanism

Double strand breakage (DSB)：

TALE binds to target sequence → FokI dimerization → cleaves DNA double strand.

Repair Path：

Non homologous terminal junction (NHEJ)Random insertion/deletion (Indels) leads to gene knockout.
Homologous Directional Restoration (HDR)External repair templates are required to achieve precise insertion or point mutation.

technological breakthrough: Shu implementationAny genomic locusAccurate cutting with a lower off target rate than early ZFN technology.

IITALENs animal modelsbuild process

（1） Standardized operating procedures

（2） Key technology optimization

step	Core Operations	innovative solution
Target design	Avoiding repetitive sequences/methylation regions, optimizing the spacer region to 14-18 bp	Software prediction (TALEN Targeted) improves success rate
Module assembly	Golden Gate cloning method：

6 modules in series (+63 architecture)
Enzymatic ligation (BsaI/BsmBI) | Built within 6 days, efficiency increased by 5 times|
|expression vector|Mammalian dual promoter vector (CMV/T7), supporting cell transfection and mRNA synthesis | adapted for multi species applications (zebrafish, mice, human iPS cells)|
|Activity verification|Yeast one hybrid system or cell reporter gene detection | Replacing time-consuming ES cell targeting, cycle shortened to 1 week|
|Embryo delivery|Microinjection of fertilized egg:
MRNA: In vitro transcription products of TALENs
RNP complex: TALE FokI protein complex | RNP reduces off target effects and enhances safety