Haploid genetic dissection of immune molecule trafficking (TECH1P.836)
| Autor: | Jingshi Shen, Eric Davis, Charles Dinarello |
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| Rok vydání: | 2014 |
| Předmět: | |
| Zdroj: | The Journal of Immunology. 192:69.4-69.4 |
| ISSN: | 1550-6606 0022-1767 |
| DOI: | 10.4049/jimmunol.192.supp.69.4 |
| Popis: | To perform their expected functions, immune molecules must be targeted to specific cellular compartments such as the plasma membrane. Improper protein targeting can lead to major forms of immune disease such as immunodeficiency and persistent inflammation. Our group developed a novel haploid genetics approach in order to unravel protein targeting on a genome-wide scale. We take advantage of human myeloid cell lines (e.g. KBM7 and HAP1) that exhibit a haploid karyotype. Retroviral insertional mutagenesis of these cells offers an efficient method to generate a library of mutant cells with complete disruption of specific genes. After selections of desired phenotypes (e.g., mislocalization of a protein), the retroviral insertion sites of the mutant clones can be mapped by deep sequencing. We have used this haploid genetics approach to characterize the plasma membrane protein CD59, a key player in the complement system. We identified ~20 genes involved in the biosynthesis, maturation, and trafficking of CD59. These genes are involved in GPI lipid synthesis, oligosaccharide modification, as well as protein trafficking at both the early and late stages of the secretory pathway. This haploid genetic screen provides the first genome-wide map of a complement protein. Many factors identified in the screen represent novel components of the complement system, and may serve as therapeutic targets for treating immune disorders. |
| Databáze: | OpenAIRE |
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