Creative Biolabs is supported by an advisory board with seasoned professionals in tumor lineage tracing. We have accumulated extensive experience from the accomplishment of different projects, during which we have received a lot of recognition and praise from our clients. Currently, we are capable of providing our clients with a variety of customized Cre-loxp recombination system-mediated lineage tracing solutions. We will accommodate the specific properties of your project and provide flexible integrated services.

Introduction to Cre-loxp Recombination System

Cre/loxP site-specific recombination system is a conditional gene knockout technology based on the interaction between Cre recombinase and loxP site. This gene editing technology makes it possible to precisely design genetic modifications, which can not only precisely introduce foreign genes, but also set genetic switches to control the expression or deletion of foreign genes in time and space, showing great application prospects.

Nowadays, Creative Biolabs has developed criteria protocols to design and synthesize Cre/loxP site-specific recombination systems. With the advantages of simple manipulation and a high recombination rate, our Cre-lox system has become a powerful tool for genetic manipulation in vitro and in vivo. Up to now, we can knock out or express a gene in a specific cell, tissue, or whole organism, or even at a specific time point, achieving spatiotemporal specific manipulation of a specific gene, which has profound implications for the study of gene function and the establishment of tumor animal models.

Fig.1 Advantages of Cre/LoxP Recombination System.

Services

AAV Vector Construction

Combining tissue-specific promoters and different AAV serotypes, Creative Biolabs offers "FLEX" systems that allow for more precise tissue-specific and timing control.

In the "FLEX" system, the target gene is located below the promoter in the reverse direction, and two "head to head" loxP are connected to each side. In the absence of Cre recombinase, the target gene could not be expressed. When the Cre recombinase is present, it can induce the "flip" of the target gene and induce its expression.

In addition, we provide a Cre recombinase-dependent trans-splicing system to establish a more efficient AAV expression vector system. To date, our dual AAV vector system can achieve a co-transfection efficiency of 25% through ITR recombination, mRNA splicing, and Cre/LoxP elimination of ITR's inhibitory effect on transcription.

Cre-LoxP Lineage Tracing Analysis

In our labs, all tumor cell groups expressing Cre are labeled with specific reporter gene proteins. The combination of specific promoter-driven Cre enzymes and reporter mice is used to produce the expression of the reporter gene. In this case, the occurrence, development, and differentiation of these tumor cells can be directly analyzed.

Fig.2 Main Services of Cre/loxP Site-Specific Recombination System.

Features of the Cre-loxP system

• High efficiency
  It can quickly trigger DNA recombination, regardless of the size and location of the target gene.
• Spatio-temporal specificity
  It can achieve gene knockout in specific kinds of organs or tissues at a specific time, avoiding the embryonic lethal effect caused by the complete gene knockout.
• Wide range of applications
  It is suitable for both eukaryotic and prokaryotic, and it can work in different tissues and under different physiological conditions.

Customize Cre-loxP System Services

• Cre element modification: It mainly focuses on the specific modification of the ligand-binding region of the estrogen receptor, so that it can only bind to certain estrogen derivatives. For example, Cre-ERT2 mutants.
• LoxP element modification: loxP elements can also be mutated in spacer and palindromic sequences, common examples are lox2272, lox511, lox5171, loxN, and loxP.
• Construction of tetO-Cre induced by tetracycline: It combines the tetracycline regulation system with the Cre-loxP system. We typically use two types of TRE-Cre and rtTA transgenic mice.

Creative Biolabs is a leading service provider that has focused on Cre-loxP system services for years. By combining cutting-edge technologies with proprietary innovations, scientists at our labs provide a full range of gene-targeting lineage tracing services. We are committed to bringing deep domain expertise to assist our clients in achieving their strategic and operational objectives. For more information, please feel free to contact us or directly send us an inquiry.

For Research Use Only | Not For Clinical Use