Homologous gene targeting in Caenorhabditis elegans by biolistic transformation.
نویسندگان
چکیده
Targeted homologous recombination is a powerful approach for genome manipulation that is widely used for gene alteration and knockouts in mouse and yeast. In Caenorhabditis elegans, several methods of target-selected mutagenesis have been implemented but none of them provides the opportunity of introducing exact predefined changes into the genome. Although anecdotal cases of homologous gene targeting in C.elegans have been reported, no practical technique of gene targeting has been developed so far. In this work we demonstrate that transformation of C.elegans by microparticle bombardment (biolistic transformation) can result in homologous recombination between introduced DNA and the chromosomal locus. We describe a scaled up version of biolistic transformation that can be used as a method for homologous gene targeting in the worm.
منابع مشابه
Reverse Genetic Strategies in Caenorhabditis elegans: Towards Controlled Manipulation of the Genome
Caenorhabditis elegans has a complete annotated genome sequence that is augmented by increasing quantities of data from high-throughput postgenomic analyses. This has led to an increasing need to identify the biological functions of specific genes using reverse genetics, i.e., moving from gene to phenotype. Fundamental to this aim is the ability to alter the structure of particular genes by mea...
متن کاملAlteration of Caenorhabditis elegans gene expression by targeted transformation.
We have produced strains carrying a synthetic fusion of parts of two vitellogenin genes, vit-2 and vit-6, integrated into the Caenorhabditis elegans genome. In most of the 63 transformant strains, the plasmid sequences are integrated at random locations in the genome. However, in two strains the transgene integrated by homologous recombination into the endogenous vit-2 gene. In both cases the r...
متن کاملGeneration of transgenic C. elegans by biolistic transformation.
The number of laboratories using the free living nematode C. elegans is rapidly growing. The popularity of this biological model is attributed to a rapid generation time and short life span, easy and inexpensive maintenance, fully sequenced genome, and array of RNAi resources and mutant animals. Additionally, analysis of the C. elegans genome revealed a great similarity between worms and higher...
متن کاملBiolistic co-transformation of rice using gold nanoparticles
ABSTRACT- In order to produce transgenic rice lines lacking selectable marker gene, biolistic co-transformation technique using gold nanoparticles was adopted. In the first step, the efficiency of different sizes of gold particles was evaluated. The results showed that the efficiency of the nanoparticles in the transformation was comparable to that of the micro particles. Subsequently, two sepa...
متن کاملTocotrienol Modulates the Expression of Proteins in Oxidative Stress-Induced Caenorhabditis Elegans
Objective: Oxidative stress that damages proteins result in aging and age related diseases. The aim of this study is to determine the effect of tocotrienol rich fraction (TRF) on the expression of proteins in oxidative stress-induced caenohabditis elegans (C.elegans) which has homologous genes to humans. Methods: The worms were treated with TRF prior to, after and continuously in separate group...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Nucleic acids research
دوره 32 4 شماره
صفحات -
تاریخ انتشار 2004