The application of viral vectors in gene therapy is rapidly developing and the therapeutic results are highly promising. However, actively and passively integrating (viral) vectors may fail to deliver, or trigger severe side effects by undirected or unintended integration into the genome of the target cell. Thus, the analysis of vector integration sites in target cells is extremely important to address both biological and safety issues. Creative Biogene, as a leading biotechnology company in the world, has extensive expertise and experience which are available to provide you with customer LAM PCR service to analyze the viral vector insertion/integration site.
LAM PCR (Linear - amplification mediated PCR) is a technology which is used for identifying and characterizing unknown flanking DNA adjacent to known DNA of any origin. More specifically, LAM-PCR has been developed to localize viral vector integration sites (IS) within the host genome [1, 2]. Genetic elements like retroviruses or transposons integrate their genome into the host genome in a (semi-) random manner [3-6]. In many cases, it is decisive to know exactly the position where these vectors integrated. With years of development, Creative Biogene has set up mature assays for several vectors, including lentiviral and most oncoviral vectors, sleeping beauty transposons, and some AAV-related vectors, etc.. Creative Biogene’s goal is to provide you with the most affordable, high-quality LAM PCR services to ensure your satisfaction in a timely and professional manner.
Analysis of the viral vector integration sites in host genomes
Localization of genetic elements and characterization of unknown flanking DNA
Elucidating biological and clinical issues
Multiarm LAM-PCR service – can detect of low copy number, multiple unknown DNA insertion site
High accuracy and sensitivity
Fast turnaround time
Creative Biogene offers custom LAM PCR services for your scientific research as follows:
Magnetic capture and double strand DNA (dsDNA) synthesis
Restriction digest and ligation of ds linker (LK)
Denaturation of synthesized dsDNA and nested PCR
Purification the PCR products and sequencing
Frequently Asked Questions of Custom LAM PCR Service
- Schmidt M, Hoffmann G, Wissler M, et al. Detection and direct genomic sequencing of multiple rare unknown flanking DNA in highly complex samples. Human gene therapy, 2001, 12(7): 743-749.
- Schmidt M, Schwarzwaelder K, Bartholomae C, et al. High-resolution insertion-site analysis by linear amplification–mediated PCR (LAM-PCR). Nature methods, 2007, 4(12): 1051-1057.
- Harkey MA, Kaul R, Jacobs MA, et al. Multiarm High-Throughput Integration Site Detection: Limitations of LAM-PCR Technology and Optimization for Clonal Analysis. Stem Cells Dev. 2007, 16(3):381-92.
- Schröder ARW, Shinn P, Chen H, et al. HIV-1 integration in the human genome favors active genes and local hotspots. Cell, 2002, 110(4): 521-529.
- Wu X, Li Y, Crise B, et al. Transcription start regions in the human genome are favored targets for MLV integration. Science, 2003, 300(5626): 1749-1751.
- Vigdal T J, Kaufman C D, Izsvák Z, et al. Common physical properties of DNA affecting target site selection of sleeping beauty and other Tc1/mariner transposable elements. Journal of molecular biology, 2002, 323(3): 441-452.