User:Joel Diaz

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Contact Info

Joel Diaz (an artistic interpretation)

I work in the Your Lab at XYZ University. I learned about OpenWetWare from Instructor, and I've joined because Required by instructor.

Education

  • 2009, BS in Biology, Loyola Marymount University

Research interests

  1. Interest 1
  2. Interest 2
  3. Interest 3

BIOL 598 - Advanced Topics in Gene Expression

Gene Therapy of Human Severe Combined Immunodeficiency (SCID)-X1 Disease [1]

Severe Combined Immunodeficiency Disorder is characterized by a block in T and NK cells. A gene therapy trial for SCID was initiated by using complementary DNA containing a defective γc Moloney retrovirus. Two infants were used as patients and after a 10 month follow up period, both T and NK cells were detectable in the patients.


Figure 1 Media:Figure1.jpeg shows results from PCR and RT-PCR tests that were run to amplify a 904 base pair stretch of γc gene sequence.

Panel A shows the results of the PCR analysis. The top portion of panel A shows the Standard Curve of a SCID patient. The standard curve was set up with a cell from a γc gene that was not infected. The second portion of panel A shows the first patient, P1. The third portion of panel A is P2, is the second patient. The samples were taken at day 150 since the start of gene therapy (d. +150). The different cells represented in each portion are the following: T CELLS ARE CD3, B CELLS ARE CD19, MONOCYTES ARE CD14, GRANULOCYTES ARE CD15, AND NK CELLS ARE CD56. Actin DNA was amplified in parallel to the γc gene as a loading control. PBMC are used as negative controls.

Panel B shows the results of the RT-PCR analysis. The same blood sample that was taken from the patients for the PCR analysis was used for RT-PCR. Actin was also amplified as a loading control.


Figure 2 Media:figure2.jpeg shows a longitudinal study of lymphocyte subsets from both patients. This set of data shows that the longer the treatment went on, the larger the increase in number of T and NK cells.


Figure 3 Media:figure3.jpeg depicts γc protein expression and lymphocyte subsets. In panel A, γc expression on B cells from patient 2 after treatment was undetectable. Panel B depicts percentages of different cells in both patients.

Publications

  1. Goldbeter A and Koshland DE Jr. An amplified sensitivity arising from covalent modification in biological systems. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6840-4. PubMed ID:6947258 | HubMed [Paper1]
  2. JACOB F and MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318-56. PubMed ID:13718526 | HubMed [Paper2]
    leave a comment about a paper here
  3. ISBN:0879697164 [Book1]
All Medline abstracts: PubMed | HubMed

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