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Wei J. Gong

Bio: Wei J. Gong is an academic researcher from University of Utah. The author has contributed to research in topics: Gene targeting & Homologous recombination. The author has an hindex of 5, co-authored 5 publications receiving 677 citations. Previous affiliations of Wei J. Gong include Stowers Institute for Medical Research.

Papers
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Journal ArticleDOI
TL;DR: It is found that ends-out targeting can be approximately as efficient as ends-in targeting, and is likely to be generally useful for Drosophila gene targeting.
Abstract: Ends-in and ends-out refer to the two arrangements of donor DNA that can be used for gene targeting Both have been used for targeted mutagenesis, but require donors of differing design Ends-out targeting is more frequently used in mice and yeast because it gives a straightforward route to replace or delete a target locus Although ends-in targeting has been successful in Drosophila, an attempt at ends-out targeting failed To test whether ends-out targeting could be used in Drosophila, we applied two strategies for ends-out gene replacement at the endogenous yellow (y) locus in Drosophila First, a mutant allele was rescued by replacement with an 8-kb y+ DNA fragment at a rate of ≈1/800 gametes Second, a wild-type gene was disrupted by the insertion of a marker gene in exon 1 at a rate of ≈1/380 gametes The I-SceI endonuclease component alone is not sufficient for targeting: the FLP recombinase is also needed to generate the extrachromosomal donor When both components are used we find that ends-out targeting can be approximately as efficient as ends-in targeting, and is likely to be generally useful for Drosophila gene targeting

374 citations

Journal ArticleDOI
01 Jan 2006-Genetics
TL;DR: It is found that Hsp70 is essential to survive a severe heat shock, but is not required to survived a milder heatshock, indicating that a significant degree of thermotolerance remains in the absence of HSp70.
Abstract: The heat-shock response is a programmed change in gene expression carried out by cells in response to environmental stress, such as heat. This response is universal and is characterized by the synthesis of a small group of conserved protein chaperones. In Drosophila melanogaster the Hsp70 chaperone dominates the profile of protein synthesis during the heat-shock response. We recently generated precise deletion alleles of the Hsp70 genes of D. melanogaster and have used those alleles to characterize the phenotypes of Hsp70-deficient flies. Flies with Hsp70 deletions have reduced thermotolerance. We find that Hsp70 is essential to survive a severe heat shock, but is not required to survive a milder heat shock, indicating that a significant degree of thermotolerance remains in the absence of Hsp70. However, flies without Hsp70 have a lengthened heat-shock response and an extended developmental delay after a non-lethal heat shock, indicating Hsp70 has an important role in recovery from stress, even at lower temperatures. Lack of Hsp70 also confers enhanced sensitivity to a temperature-sensitive lethal mutation and to the neurodegenerative effects produced by expression of a human polyglutamine disease protein.

129 citations

Journal ArticleDOI
01 Nov 2004-Genetics
TL;DR: The results show that genomic deletions of varied sizes can be readily generated by homologous recombination in Drosophila, including one deletion that spanned ∼47 kb.
Abstract: Homologous recombination can produce directed mutations in the genomes of a number of model organisms, including Drosophila melanogaster. One of the most useful applications has been to delete target genes to generate null alleles. In Drosophila, specific gene deletions have not yet been produced by this method. To test whether such deletions could be produced by homologous recombination in D. melanogaster we set out to delete the Hsp70 genes. Six nearly identical copies of this gene, encoding the major heat-shock protein in Drosophila, are found at two separate but closely linked loci. This arrangement has thwarted standard genetic approaches to generate an Hsp70-null fly, making this an ideal test of gene targeting. In this study, ends-out targeting was used to generate specific deletions of all Hsp70 genes, including one deletion that spanned ∼47 kb. The Hsp70-null flies are viable and fertile. The results show that genomic deletions of varied sizes can be readily generated by homologous recombination in Drosophila.

115 citations

Book ChapterDOI
TL;DR: Two methods of gene targeting in Drosophila are presented, identical in concept to gene replacement techniques used routinely in mammalian and yeast cells, but each method has separate strengths and drawbacks.
Abstract: We present detailed protocols for two methods of gene targeting in Drosophila. The first, ends-out targeting, is identical in concept to gene replacement techniques used routinely in mammalian and yeast cells. In Drosophila, the targeted gene is replaced by the marker gene white + (although options exist to generate unmarked targeted alleles). This approach is simple in both the molecular cloning and the genetic manipulations. Ends-out will likely serve most investigators' purposes to generate simple gene deletions or reporter gene "knock-ins." The second method, ends-in targeting, targets a wild-type gene with an engineered mutated copy and generates a duplication structure at the target locus. This duplication can subsequently be reduced to one copy, removing the wild-type gene and leaving only the introduced mutation. Although more complicated in the cloning and genetic manipulations (see Note 1), this approach has the benefit that the mutations may be introduced with no other remnant of the targeting procedure. This "surgical" approach will appeal to investigators who desire minimal perturbation to the genome, such as single nucleotide mutation. Although both approaches appear to be approximately equally efficient (see Note 2), each method has separate strengths and drawbacks. The choice of which approach is best depends on the researcher's goal.

74 citations

Patent
09 May 2003
TL;DR: In this paper, the endonuclease and recombinase enzymes are used so that homologous recombination occurs between the DNA segment of the donor construct and a selected gene of the host organism, thus forming a host with containing the recombinogenic donor.
Abstract: A gene targeting method for use in a host organism whereby the host organism is transfected with an ends-out donor construct. Further transfecting the host organism with two transgenes expressing endonuclease and recombinase enzymes. The endonuclease and recombinase enzymes are used so that homologous recombination occurs between the DNA segment of the donor construct and a selected gene of the host organism, thus forming a host with containing the recombinogenic donor. The progeny of the host organism, which include the recombinogenic donor are then selected.

11 citations


Cited by
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Journal ArticleDOI
12 Aug 2005-Cell
TL;DR: Yorkie (Yki), the Drosophila ortholog of the mammalian transcriptional coactivator yes-associated protein (YAP), is identified as a missing link between Wts and transcriptional regulation and is a critical target of the Wts/Lats protein kinase and a potential oncogene.

1,621 citations

Journal ArticleDOI
02 Sep 2004-Neuron
TL;DR: The results support the second model of Or83b function, which encodes an atypical odorant receptor that plays an essential general role in olfaction and disrupts behavioral and electrophysiological responses to many odorants.

1,221 citations

Journal ArticleDOI
01 Jan 2013-Genetics
TL;DR: A bacterial CRISPR RNA/Cas9 system is adapted to precisely engineer the Drosophila genome and it is reported that Cas9-mediated genomic modifications are efficiently transmitted through the germline.
Abstract: We have adapted a bacterial CRISPR RNA/Cas9 system to precisely engineer the Drosophila genome and report that Cas9-mediated genomic modifications are efficiently transmitted through the germline. This RNA-guided Cas9 system can be rapidly programmed to generate targeted alleles for probing gene function in Drosophila.

1,067 citations

Journal ArticleDOI
02 May 2003-Science
TL;DR: A general method for improving the efficiency of gene targeting would be valuable in many circumstances, as would the extension of this genetic tool to a wider range of applications.
Abstract: Gene targeting—the process of gene replacement by homologous recombination—is a very useful but typically inefficient technique ( [1][1] ). A general method for improving the efficiency of gene targeting would be valuable in many circumstances, as would the extension of this genetic tool to a

831 citations

Journal ArticleDOI
TL;DR: It is shown that injection of RNA into the Drosophila embryo can induce highly efficient mutagenesis of desired target genes in up to 88% of injected flies.

824 citations