The Human Insulin Gene Displays Transcriptionally Active Epigenetic Marks in Islet-Derived Mesenchymal Precursor Cells in the Absence of Insulin Expression

Stem Cells ◽  
2007 ◽  
Vol 25 (12) ◽  
pp. 3223-3233 ◽  
Author(s):  
Vesco Mutskov ◽  
Bruce M. Raaka ◽  
Gary Felsenfeld ◽  
Marvin C. Gershengorn
Keyword(s):  
Human Insulin ◽  
Precursor Cells ◽  
Insulin Gene ◽  
Epigenetic Marks ◽  
Insulin Expression ◽  
Human Insulin Gene ◽  
Mesenchymal Precursor Cells ◽  
Mesenchymal Precursor

scholarly journals Modification of a Constitutive to Glucose-Responsive Liver-Specific Promoter Resulted in Increased Efficacy of Adeno-Associated Virus Serotype 8-Insulin Gene Therapy of Diabetic Mice

Cells ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 2474
Author(s):  
Kian Chuan Sia ◽  
Zhen Ying Fu ◽  
Roy Y. Calne ◽  
Amit C. Nathwani ◽  
Kok Onn Lee ◽  
...  
Keyword(s):  
Human Insulin ◽  
Liver Toxicity ◽  
Insulin Gene ◽  
Diabetic Mice ◽  
Insulin Expression ◽  
Human Insulin Gene ◽  
Vector Genome ◽  
Virus Serotype

We have previously used a hepatotropic adeno-associated viral (AAV) vector with a modified human insulin gene to treat diabetic mice. The HLP (hybrid liver-specific promoter) used was constitutively active and non-responsive to glucose. In this study, we examined the effects of addition of glucose responsive elements (R3G) and incorporation of a 3′ albumin enhancer (3′iALB) on insulin expression. In comparison with the original promoter, glucose responsiveness was only observed in the modified promoters in vitro with a 36 h lag time before the peak expression. A 50% decrease in the number of viral particles at 5 × 109 vector genome (vg)/mouse was required by AAV8-R3GHLP-hINSco to reduce the blood sugar level to near normoglycemia when compared to the original AAV8-HLP-hINSco that needed 1 × 1010 vg/mouse. The further inclusion of an 860 base-pairs 3′iALB enhancer component in the 3′ untranslated region increased the in vitro gene expression significantly but this increase was not observed when the packaged virus was systemically injected in vivo. The addition of R3G to the HLP promoter in the AAV8-human insulin vector increased the insulin expression and secretion, thereby lowering the required dosage for basal insulin treatment. This in turn reduces the risk of liver toxicity and cost of vector production.

Lack of association of the polymorphic locus in the 5'-flanking region of the human insulin gene and diabetes in American blacks

Diabetes ◽  
1985 ◽  
Vol 34 (5) ◽  
pp. 433-439 ◽  
Author(s):  
S. Elbein ◽  
P. Rotwein ◽  
M. A. Permutt ◽  
G. I. Bell ◽  
N. Sanz ◽  
...  
Keyword(s):  
Human Insulin ◽  
Polymorphic Locus ◽  
Insulin Gene ◽  
Human Insulin Gene ◽  
Flanking Region ◽  
American Blacks

Cloning and Expression of Recombinant Human Insulin Gene in Pichia pastoris

2019 ◽  
Vol 10 (01) ◽  
Author(s):  
Rafid A. Abdulkareem
Keyword(s):  
Pichia Pastoris ◽  
Human Insulin ◽  
Expression Vector ◽  
Expression System ◽  
Insulin Gene ◽  
Cloning And Expression ◽  
Pcr Analysis ◽  
Major Band ◽  
Human Insulin Gene ◽  
Pichia Pastoris Expression System

The main goal of the current study was cloning and expression of the human insulin gene in Pichia pastoris expression system, using genetic engineering techniques and its treatment application. Total RNA was purified from fresh normal human pancreatic tissue. RNA of good quality was chosen to obtain a first single strand cDNA. Human preproinsulin gene was amplified from cDNA strand, by using two sets of specific primers contain EcoR1 and Notl restriction sites. The amplified preproinsulin gene fragment was double digested with EcoRI and Not 1 restriction enzymes, then inserted into pPIC9K expression vector. The new pPIC9K-hpi constructive expression vector was transformed by the heat-shock method into the E.coli DH5α competent cells. pPic9k –hpi, which was propagated in the positive transformant E. coli cells, was isolated from cells and then linearised by restriction enzyme SalI, then transformed into Pichia pastoris GS115 using electroporation method. Genomic DNA of His+ transformants cell was extracted and used as a template for PCR analysis. The results showed, that the pPic9k – hpi was successfully integrated into the P. pastoris genome, for selected His+ transformants clones on the anticipated band at 330 bp, which is corresponded to the theoretical molecular size of the human insulin gene. To follow the insulin expression in transformans, Tricine–SDS gel electrophoresis and Western blot analysis were conducted. The results showed a successful expression of recombinant protein was detected by the presence of a single major band with about (5.8 KDa) on the gel. These bands correspond well with the size of human insulin with the theoretical molecular weight (5.8 KDa).

scholarly journals Variation in the sequence and modification state of the human insulin gene flanking regions

1982 ◽  
Vol 10 (7) ◽  
pp. 2225-2240 ◽  
Author(s):  
Axel Ullrich ◽  
Thomas J. Dull ◽  
Alane Gray ◽  
John A. Philips ◽  
Stephan Peter
Keyword(s):  
Human Insulin ◽  
Insulin Gene ◽  
Human Insulin Gene ◽  
Flanking Regions

Polymorphism in the 5'-flanking region of the human insulin gene and its possible relation to type 2 diabetes

Science ◽  
1981 ◽  
Vol 213 (4512) ◽  
pp. 1117-1120 ◽  
Author(s):  
P Rotwein ◽  
R Chyn ◽  
J Chirgwin ◽  
B Cordell ◽  
H. Goodman ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Human Insulin ◽  
Insulin Gene ◽  
Human Insulin Gene ◽  
Flanking Region

A Comparison of Mesenchymal Precursor Cells and Amnion Epithelial Cells for Enhancing Cervical Interbody Fusion in an Ovine Model

Neurosurgery ◽  
2011 ◽  
Vol 68 (4) ◽  
pp. 1025-1035 ◽  
Author(s):  
Tony Goldschlager ◽  
Peter Ghosh ◽  
Andrew Zannettino ◽  
Mark Williamson ◽  
Jeffrey Victor Rosenfeld ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Interbody Fusion ◽  
Precursor Cells ◽  
Ovine Model ◽  
Mesenchymal Precursor Cells ◽  
Mesenchymal Precursor ◽  
Amnion Epithelial Cells
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