Overview
Get rid of selection cassettes and other unwanted sequences in your genome engineering projects
With the Cre-LoxP site-specific recombination system used in most of our HR Targeting Vectors, you can get rid of extra sequences, such as selection cassettes, by transiently expressing the Cre recombinase from the pCMV-CRE Cre Recombinase Expression Plasmid.
How It Works
Removing a selection cassette with Cre recombinase
Cre recombinase catalyzes site-specific recombination between two LoxP sites. After recombination occurs, the sequences between the two LoxP sites are excised from the genome and a single LoxP site remains (Figure 1). Thus, with our HR Targeting Vectors that contain two LoxP sites, all the vector sequences that lie between the LoxP sites can be removed through transient expression of the Cre recombinase (Figure 1).
Figure 1. Removing a selection cassette using Cre-LoxP. The example shown demonstrates Cre-LoxP excision during genome editing but applies to any application where regions of an HR Targeting Vector with LoxP sites are integrated into the genome. In Step 1, Cas9 activity leads to a double-stranded break (DSB) in the genome. In Step 2, the regions of an HR Targeting Vector (also called an HR donor vector) that lie between the two homologous arms become integrated into the genome via homologous recombination. This integrated region typically contains a selection cassette with flanking LoxP sites to simplify finding clones where genome editing has occurred. In Step 3, the selection cassette is removed by transient expression of Cre recombinase.
Resources
System Biosciences,简称SBI，美国加州湾区新成立的生技公司，致力于独特、创新生物技术之开发，以研发利于基因及蛋白质功能鉴定、研究之崭新方法和工具为宗旨。 现阶段研发重心为RNA干扰（RNAi）研究之相关工具。 System Biosciences (SBI) 致力于开发独特、革新的技术，为客户研究蛋白组学和基因组学功能提供研究工具。SBI 是专业的慢病毒产品公司，提供基于慢病毒的所有相关产品、质粒、试剂盒及相关配套试剂和慢病毒延伸产品如IPS细胞多功能性诱导试剂盒和RNAi筛选文库。
SBI focuses on developing unique, innovative technologies that provide researchers with the tools to investigate and understand genomic and proteomic function. Our mission is to provide tools for the genome-wide analysis of the mechanisms that regulate cellular processes and biological responses.
Headquartered in Palo Alto about 30 miles south of San Francisco, SBI is geographically surrounded by highly successful research biotech companies (e.g., Affymetrix, Life Technologies, and Genentech) and some of the premier life science institutes in the world, including Stanford and the University of California, San Francisco.
As a small private biotechnology company, SBI capitalizes on a rich network of consultants and colleagues and continually collaborates to accelerate its development of innovative and novel applications. Currently, SBI has partnerships with over thirty scientists from several institutes.System Biosciences (SBI) consists of a highly motivated team committed to realizing the potential of its mission to develop and bring to market unique and innovative technology to investigate and understand genomic and proteomic function.
Possessing a diverse range of experience and knowledge, SBI's management and staff bring the skills, talent, and interest needed to support the company's continuing rapid growth.
A strong and proven background in creative research and product development with proficiency in genetic analysis, microarray technology, and cell biology
Extensive business development and management experience
A keen understanding of the needs of life scientists
Expertise in developing, marketing, and supporting consistent, high-quality research products
A clear focus on the need to build and maintain lasting customer relationships through support, service, and close customer interaction
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