Molecular Medicine
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RNAi ---> siRNA ---> RNAa ---> saRNA --->ASO --->miRNA ---> RNA/DNA synthesis ---> new bio-farmaceutics based on nucleic acid molecule --->  

DNA/RNA analysis ---> gDNA ---> mtDNA ---> SNP ---> STR ---> sequencing ---> epigenetics ---> microarray --->  laboratory diagnostics --->

Engineering recombinant-protein --->  genetic constructs ---> protein synthesis & purification ---> bio-farmaceutics ---> 

   

Genetic Laboratory of BioTe21 headed by Adam Master would like to present some original methods & technologies, that may be used in:
 
<> Gene expression regulation and gene therapy (to find more, please click the link: gene enhancement.pdf),

<> Medical laboratory diagnostics (to find more, please click the link: cancer diagnostics.pdf).

<> Genetic identification (to find more, please click the link: mtDNA identification.pdf)

Technologies designed and elaborated by Adam Master:
 


Medical laboratory diagnostics

Genetic Laboratory of BioTe21 headed by Adam Master would like to present some new laboratory diagnostics allowing fast and routine screening for:

  •  Increased risk of developing UV-dependent melanoma malignum (MC1R, CDKN2A),

  •  Hereditary Prostate, Breast and Ovarian Cancer (BRCA1, BRCA2, CHEK2, NBS1),

  •  Factor V Leiden (SNP: R506Q; F5) prothrombin (SNP: G20210A; F2),

  • and many others (based on SNP/mutation detection).


BioTe21 Adam Master in GeneTests of NCBI:


 
 

melTest kit®  is a unique product used in UV-dependent skin cancer (malignant melanoma) predisposition diagnosis. MelTest® has been designed thanks to many years of experience of BioTe21 in the field of genetic diagnostics. This experience allowed us to create a test specifically designed to detect predisposition for UV-dependent skin cancer. The diagnostics of UV-dependent skin cancer is based on DNA sequencing technology, as well as fluorescent analysis of SNP (mini-sequencing). An analysis of up to 15 SNP-type mutations in MC1R gene and up to 6 mutations in CDKN2A gene (both in correlation with high risk of skin cancer occurrence) is performed.

Using our technology, there is no need for special preparation or adaptation of the laboratory. Analysis may be performed in any genetic laboratory (equipped with a thermocycler and a sequenator) an is an excellent way to make your offer of services even more attractive – which may be especially interesting for companies specialized in paternity testing using STP (Short Tandem Repeats) analysis of genomic DNA.

Understanding the information about mutations associated with some hereditary diseases, and using preventive genetic diagnostics, may help the individuals in changing their life-styles (especially risky behavior, like an extensive sun-light exposition) if they have UV-dependent skin cancer predisposition, and therefore, save their lives.


Hereditary Breast and Ovarian Cancer.
BioTe21 Adam Master has also prepared and introduced a number of new technologies in the field of molecular diagnostic medicine -  unique tests for hereditary cancer predisposition including Hereditary Prostate, Breast and Ovarian Cancer (BRCA1, BRCA2, CHEK2, NBS1).


 Detection of BRCA1 mutations, including: 5382insC, T300G, 4153delA, 3819del5, G3867T, 185dAG, T309C, T74381G (Met1775Arg), IVS20+60ins12, 3875del4, IVS14+1G>A, 5370C/T, 4182delAATC, 794delT, 5149del4,

 BRCA2: Lys3326X, Tyr3401Met,Tyr3401Met, G8410A (Val2728Ile), C5972T (Thr1915Met), 4088delA (ter1291), G7985A (Trp2586Ter), 488delCT, G4486T, A1093TC,

 CHEK2: 1100delC, I157T, IVS2+1G>A, del5395, Glu239Lys (G715A), Glu239Stop (G715T),

 NBS1: C643T, R215W, 657del5.

 


Diagnostics of Factor V Leiden
(SNP: R506Q; F5) prothrombin (SNP: G20210A; F2): 

 
 
             

 

Enhancement of protein synthesis

eRNA– enhancing RNA– selective enhancing of protein synthesis. The technology complements genes silencing methods (siRNA, miRNA). However, what makes eRNA different and exceptional, is the fact, that instead of inhibiting, it enhances the translational efficiency of chosen genes in selective manner.

Both in biology and molecular medicine, selective silencing of genes is already being performed by the means of: 1) synthetic siRNA or antisense molecules as well as 2) non-selective, native cellular miRNAs. However, until now, there was no method for selective enhancement of chosen genes. Just like siRNA (artificial small inhibitory RNA) and miRNA (naturally occurring inhibitory microRNA), the eRNA may use RNAi related phenomena and/or RNAse H pathways to induce cellular response. However, what makes the eRNA different and exceptional, is the fact, that instead of inhibiting it leads to protein synthesis enhancement of selected genes or gene fragments even by 500%, in selective manner.
 

 


Some properties of the eRNA:
• works as a trans-acting factor binding to a cis-acting element,
• acts as a protein synthesis enhancer at the level of translational control,
• does not change in significant way the level of transcription,
• targets a selected mRNA - as designed before on the basis of a chosen sequence of a single gene or even its allele characterized by a single nucleotide polymorphism (SNP).


Some practical uses for eRNAs:
1. enhancement of suppressors, mutators, modulators and other genes involved
 in anti-cancer response: (TP53, BAX, CDKN2A, CDK4, RB1, NOD2, CHEK2,
INBS1, XPD, MC1R, MSH2, MLH1, MSH6, BRCA1, BRCA2, APC and others),
2. enhancement of anti-viral response  by activation of: interferons (IFNA1,
IFNA2, IFNB, IFNG)  and others,
3. enhancement of anti-aging genes such as mutator genes, telomerase (TERT),
 collagens: (COL1A1-COL9A1) and others that activation maybe useful in
aesthetic medicine,
4. enhancement of genes involved in hereditary diseases,
5. enhancement of genes, that expression is too low or is wanted to be higher,
including cytokines and growth factors (Il1-Il10, VEGF, TGF, TNF, EGF, FGF), trans-membrane receptors, protein hormones (INS, GH1, IGH), proteins of signaling pathways, apoptotic pathways, enzymes and other genes important for genetic studies and industrial biotechnology,
6. enhancement of chosen splicing variants of a single gene (for example: the longer variant of the BRCA1 gene),
7. selective enhancing of a correct allele of  a gene in heterozygous cells (SNP-differing gene alleles wherein one of the alleles is mutated, the other one is correct),
8. overexpression of a selected gene at the level of translation,
9. studies on cell cycle, proliferation, growth, differentiation, embryogenesis, carcinogenesis and apoptosis, endocytosis, exocytosis, internalization and intercellular communication,
10. studies on interaction between proteins,
11. enhancing translational efficiency in technological processes of recombined proteins,
12. and many more that we can you propose you on the basis of your studies.



 
Genetic identification

BioTe21 Genetic Laboratory presented also an optimized technology of mitochondrial DNA analysis and its implementation. The unique mtDNAtest kit was designed for the purpose of genetic identification and mtDNA genotype determination. The kit is useful in population genetics, in the field of determining haplotypes and haplogroups helpful in lineage analysis and family tree construction.


Many years of professional experience in the field of forensic molecular medicine, as well as the work with mtDNA allowed me to design and introduce a new test, which is, in technical terms, an extremely easy way to differentiate SNPs (Single Nucleotide Polymorphism) in mtDNA, based on the method of fluorescent DNA fragments separation (mtDNAtest-SNP module). mtDNAtest kit will allow to yield reliable results, without the need of difficult sequence analysis. Specially chosen set of analyzed SNP allows analyzing the most hyper variable HVI and HVII sequences’ locations, therefore it is useful especially in genetic identification and comparative mtDNA analysis of biological traces.

Apart from the mentioned innovative test of specific SNPs, it can be used a standard kit for the variable mtDNA regions’ sequencing with specially chosen set of oligonucleotides, allowing to yield highest quality results every single time (mtDNAtest-SEK module).

MtDNA analysis, with the help of the technologies, does not require a specially equipped lab and, therefore, may be performed in any genetic laboratory routinely analyzing STR (Short Tandem Repeats) of genomic DNA (gDNA). All you need is a thermocycler and a standard sequenator. The above presented mtDNAtest kit’s modules use different basic methods (minisequencing or classic sequencing) and may be employed separately.

VALIDATION:

 


 
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