[52] Gene isolation by retroviral tagging

1987 ◽  
pp. 469-481 ◽  
Author(s):  
Stephen P. Goff
Keyword(s):  
Gene Isolation

Exosomes: Structure, Biogenesis, Types and Application in Diagnosis and Gene and Drug Delivery

2020 ◽  
Vol 20 (3) ◽  
pp. 195-206 ◽  
Author(s):  
Shriya Agarwal ◽  
Vinayak Agarwal ◽  
Mugdha Agarwal ◽  
Manisha Singh
Keyword(s):  
Drug Delivery ◽  
Gene Therapy ◽  
Immune Responses ◽  
Biological Membrane ◽  
Gene Isolation ◽  
Delivery Vehicle ◽  
Scientific Communities ◽  
Therapeutic Regime ◽  
Targeted Gene Therapy ◽  
Delivery Mechanism

Abstract: In recent times, several approaches for targeted gene therapy (GT) had been studied. However, the emergence of extracellular vesicles (EVs) as a shuttle carrying genetic information between cells has gained a lot of interest in scientific communities. Owing to their higher capabilities in dealing with short sequences of nucleic acid (mRNA, miRNA), proteins, recombinant proteins, exosomes, the most popular form of EVs are viewed as reliable biological therapeutic conveyers. They have natural access through every biological membrane and can be employed for site-specific and efficient drug delivery without eliciting any immune responses hence, qualifying as an ideal delivery vehicle. Also, there are many research studies conducted in the last few decades on using exosome-mediated gene therapy into developing an effective therapy with the concept of a higher degree of precision in gene isolation, purification and delivery mechanism loading, delivery and targeting protocols. This review discusses several facets that contribute towards developing an efficient therapeutic regime for gene therapy, highlighting limitations and drawbacks associated with current GT and suggested therapeutic regimes.

Long-range genomic map of the Duchenne muscular dystrophy (DMD) gene: Isolation and use of J66 (DXS268), a distal intragenic marker

Genomics ◽  
1987 ◽  
Vol 1 (4) ◽  
pp. 329-336 ◽  
Author(s):  
G.J.B. van Ommen ◽  
C. Bertelson ◽  
H.B. Ginjaar ◽  
J.T den Dunnen ◽  
E. Bakker ◽  
...  
Keyword(s):  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Long Range ◽  
Gene Isolation ◽  
Dmd Gene ◽  
Genomic Map

scholarly journals Isolation of Human Transcripts Expressed in Hamster Cells from YACs by cDNA Representational Difference Analysis

1999 ◽  
Vol 9 (2) ◽  
pp. 182-188
Author(s):  
Jessie Gu ◽  
Xin-Yuan Guan ◽  
Melissa A. Ashlock
Keyword(s):  
Cell Line ◽  
Mammalian Cells ◽  
Positional Cloning ◽  
Yeast Artificial Chromosome ◽  
Expressed Sequence Tag ◽  
Gene Isolation ◽  
Representational Difference Analysis ◽  
Difference Analysis ◽  
Artificial Chromosomes ◽  
Link Type

Gene isolation methods used during positional cloning rely on physical contigs consisting of bacterial artificial chromosomes, P1, or cosmid clones. However, in most instances, the initial framework for physical mapping consists of contigs of yeast artificial chromosome (YACs), large vectors that are suboptimal substrates for gene isolation. Here we report a strategy to identify gene sequences contained within a YAC by using cDNA representational difference analysis (RDA) to directly isolate transcripts expressed from the YAC in mammalian cells. The RDA tester cDNAs were generated from a previously reported hamster cell line derived by stable transfer of a 590-kb YAC (911D5) that expressed NPC1, the human gene responsible for Niemann-Pick type C (NP-C). The driver cDNAs were generated from a control hamster cell line that did not contain the YAC that expressed NPC1. Among the gene fragments obtained by RDA,NPC1 was the most abundant product. In addition, two non-NPC1 fragments were isolated that were mapped to and expressed from 911D5. One of these RDA gene fragments (7-R) spans more than one exon and has 98% sequence identity with a human cDNA clone reported previously as an expressed sequence tag (EST), but not mapped to a chromosomal region. The other fragment (2-R) that had no significant sequence similarities with known mammalian genes or ESTs, was further localized to the region of overlap between YACs911D5 and 844E3. The latter YAC is part of a contig across the NP-C candidate region, but does not contain NPC1. This two-part approach in which stable YAC transfer is followed by cDNA RDA should be a useful adjunct strategy to expedite the cloning of human genes when a YAC contig is available across a candidate interval.[The sequence data described in this paper have been submitted to GenBank under accession nos. AF117641 and AF117642.]

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