The popularity of genetically modified rodents as models for the studies of gene function, modeling of human disease to either understand disease mechanisms or to aid drug development, and xenotransplantation in recent years has been accompanied by the development of numerous tools for genome manipulation. These tools include various transgenic approaches, homologous recombination-based gene targeting technologies, the latest nuclease-mediated genome editing techniques, and so on. With these technologies, Creative Biolabs is capable of providing the most comprehensive model creation services to our clients around the world.
DNA microinjection is one of the most commonly used methods of transferring or introducing novel exogenous genes into animals. The gene of interest can be randomly inserted into the genome of a newly fertilized egg via a micromanipulator under the microscope. Besides, microinjection techniques are also used for ES cell injection and CRISPR injection in the ESC-mediated gene targeting process and the CRISPR/Cas9-mediated gene editing, respectively.
Lentivectors are used to generate transgenic animal models via the transduction of ES cells or early embryos. This method has exhibited advantages such as stable expression of the transgenic DNA and high efficiency. Summarily, three principles of successful lentiviral transgenesis are design and generation of the optimal lentiviral vector, production of high titer lentivirus, and skillful manipulation of microinjection.
Transposon can be used as gene transfer vectors to generate transgenic animals with high efficiency. They have shown unique advantages compared to other genetic manipulation techniques. At Creative Biolabs, transposons such as Sleeping Beauty, Tol2, and PiggyBac are available to create the ideal transgenic animal models you've desired.
Fig.1 Targeted genome editing tools.
(Cheng et al. 2014)
Gene targeting in embryonic stem cells based on homologous recombination is a technology used to specifically integrate an exogenous gene into a certain locus in the target cell genome. This technique is used to create models that lack specific genes (“Knock-in”) or those in which one existing gene has been replaced by another that has been engineered (“Knock-out”).
In the knock-out category, the target gene can be permanently inactivated in every cell of the organs (Conventional Knock-out Models) or can be specifically deleted at particular time points (Inducible Knock-out Models), in particular tissues (Tissue-specific Knock-out Models). Besides, a multipurpose mouse model (KO First Models) in which both conventional and conditional knock-out models can be generated simultaneously.
In the knock-in category, the homologous recombination-mediated approach allows the site-specific insertion of a foreign DNA, including a reporter gene (e.g., EGFP, mRFP, mCherry and LacZ) and functional cDNA (e.g., Cre and Dre). Further, fixed type knock-in (e.g., ROSA26 Knock-in Models) is also covered.
Nuclease-Mediated Genome Editing
Due to the low efficient, time-consuming and costly characteristics of homologous-mediated targeting strategy, nuclease-based genome editing approaches, including CRISPR/Cas9 and TALEN, have been greatly promoted in recent years. These two systems offer a more convenient and much simpler method for genome editing, thus playing a significant role in multiple fields.
Although CRISPR/Cas9 is a newer and more popular technology, reported to be a major breakthrough in genome editing, TALEN still remains to be advantageous in certain respects. As a result, at Creative Biolabs, both CRISPR/Cas9 and TALEN-mediated genome editing technologies are available for our clients to choose from, depending on their specific demands and research objectives. Please click the links below for more information:
See below for more information about the related sections for genetically model creation:
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