what is genetic engineering?
variety of techniques for the purposeful manipulation of genetic material to enhance, alter and repair the function of the cell.
what is criper cas 9
CRISPR/Cas9 edits genes by precisely cutting DNA and then letting natural DNA repair processes to take over.
what is car-t cells
Car t cells is a type of immunotherapy that modifies a patient’s immune system to allow it to be more effective in finding and destroying diseased cells.
what is the methodology process of car -t cells
- Sample of patient’s peripheral blood mononuclear cell separating the t cells from the white blood.
- T-cells are isolated and cultured and inserted with the CAR gene via retroviral vectors.
- The new cell is expanded ex vivo to high numbers and infused back into the patient to elicit a durable anti-tumour response.
The Process of Car-T Cells to treat Cancer ?
- CAR is made up of a single-chain antibody fragment in which identifies CD19 (protein used to diagnose cancers)
- The Car t cell expresses a protein on the surface which binds to the cancerous cells.
- The cells multiply + signal to parts of the immune system - cytokines to come to the site of the cancer cell
- causing significant inflammation focused on the cancer cell
what cancers are being treated
- diffuse large B-cell lymphoma
- follicular lymphoma
mantle cell lymphoma
-multiple myeloma - B-cell acute lymphoblastic leukemia
use on solid tumours
currently clinical trails are being assessed
3 in 11 patients who previously had active disease of of paediatric neuroblastoma before the trial achieved a complete remission
use on Lymphatic system
Phase II clinical trials were presented to Yescarta The results illustrated substantial improvements, contrasted to standard care with 83% of the CAR T-treated patients positively responding to the therapy, compared to 50% who positively responded to the standard second-line treatment.
Issues, Limitations and Conflicting Views
use of vectors
- Lentiviral vectors could result in insertional mutagenesis which has potential to alter the expression of the CAR gene,
Long Term Impacts on Patients
- potentially severe toxicities with serious adverse events such as cytokine release syndrome
a systemic inflammatory response caused by cytokines released by infused CAR T cells can lead to widespread reversible organ dysfunction
Future Directions for Car-T Cell
the scope to further optimize Car-T cell design and delivery to cure thousands of people, is still underway.
completed on the use of lentiviral vector to transfer the Car gene to ensure low risk of insertional of mutagenesis
For the future this method needs to undergo more trials on blood cancers, and all with solid cancers to make it more effective to allow majority to of patient’s cancer go into remission.
what is genetic transformation
process of integrating exogenous DNA into the germ-line of whole organisms so that it is inherited in subsequent generations.
why are bacteria useful for genetically transformed
reproduces quickly - fast reproduction of offspring will allow you to identify and assess the new traits in the next generation
singled cell meaning only one cell is modified.
what are the three genes in plasmid
- ARAC: the protein in the presence of Arabinose caused the transcription of the GTP gene (switches on and off)
- GFP: the gene for a fluorescent
- bete-lactamase gene allows the transformed bacteria to be resistant to ampliliin
what is a plasmid
small circular DNA molecules separated from cells chromosomes DNA
what was the purpose for Cacl2 transformation solution
shields negative charge on dna
Purpose of pre heat shock (in ice)
slows fluid plasma membrane for greater shock
