Sickle Cell - Targeted Gene Repair - Advanced Progress
Notes:
(1) This study was performed at the University of Texas Medical branch and NOT the University of Deleware. The significance of this is that the doctor cited in the
study as Kmiec E MIGHT BE a different doctor than the Kmiec EB who works at NPRO.
(2) Another doctor cited is Davis BR (in my database)
and his background is in the area of Small Fragment DNA (SFHR) - however - this article is NOT about SFHR but straight-forward Chimeric oligonucleotides. So I am
not certain whether their success was due to
straight-forward Chimeric oligonucleotides or - in some
part - Small Fragment DNA.
comment -
this shit sounds like the real deal ...
this article is EASY to READ ...
they are using HUMAN cells .....
1: Gene Ther 2002 Jan ;9(2):118-26 Related Articles, Links
Targeted beta-globin gene conversion in human hematopoietic CD34(+ )and Lin(-)CD38(-)cells.
Liu H, Agarwal S, Kmiec E, Davis BR.
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA.
Chimeric oligonucleotides have been used successfully to correct point and frameshift mutations in several cell types, as well as in animal and plant models.
However, their application to primitive human blood cells has been limited.
In this investigation, chimeric oligonucleotides designed to direct a site-specific nucleotide exchange in the human beta-globin gene (an A to T substitution within codon 6) were introduced into normal human CD34(+) and Lin(-)CD38(-) cells via microinjection.
This A to T nucleotide exchange introduces the single site mutation responsible for sickle cell anemia. In 23% of experimental samples, gene conversion was detected in the progeny of microinjected CD34(+) and Lin(-)CD38(-) cells that were cultured for at least 4 weeks.
In addition, gene conversion was detected in the erythroid progeny of Lin(-)CD38(-) cells at the mRNA level.
Conversion rates as high as 10-15% in 11% (five of 44) of experimental samples were confirmed by allele-specific PCR and sequence analysis of genomic DNA from the progeny of microinjected Lin(-)CD38(-) cells.
Given that as few as 10% normal hematopoietic cells are sufficient to keep patients free of sickle cell disease, the level of conversion we have achieved in some samples may well be of therapeutic benefit in patients with sickle cell disease.
PMID: 11857070 [PubMed - indexed for MEDLINE]
ncbi.nlm.nih.gov
Additional Information:
below is the 1996 article by Kmiec EB about Sickle Cell ...
1: Science 1996 Sep 6 ;273(5280):1386-9 Related Articles, OMIM, Cited in PMC, Books, LinkOut
Science. 1997 Mar 7;275(5305):1404-5.
Correction of the mutation responsible for sickle cell anemia by an RNA-DNA oligonucleotide.
Cole-Strauss A, Yoon K, Xiang Y, Byrne BC, Rice MC, Gryn J, Holloman WK, Kmiec EB.
Department of Pharmacology, Kimmel Cancer Center, Thomas Jefferson University, 233 South 10th Street, Philadelphia, PA 19107, USA.
A chimeric oligonucleotide composed of DNA and modified RNA residues was used to direct correction of the mutation in the hemoglobin betaS allele. After introduction of the chimeric molecule into lymphoblastoid cells homozygous for the betaS mutation, there was a detectable level of gene conversion of the mutant allele to the normal sequence. The efficient and specific conversion directed by chimeric molecules may hold promise as a therapeutic method for the treatment of genetic diseases.
ncbi.nlm.nih.gov
John McCarthy |
