Gene transfer and Expression Literature Papers

Anonymous
timer Asked: Jun 20th, 2018
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Question description

you must complete 4 separate papers. Select the papers from the current peer-reviewed original research literature (nothing before 2016 is acceptable) that uses gene transfer and expression as its major goal (it can be the only goal but doesn’t have to be) in journals like Science, Nature, Cell, Molecular and Cellular Biology, etc. Trade magazines, like BioTechniques, are NOT acceptable for this.

Written paper must include

1. Overall scientific goal(s)

2. Gene being transferred, how and why

3. Vector specifics (such as plasmid, regulatory cassettes, etc)

4. Recipient specifics (what cells and why)

5. Detection of transfer/expression

6. Your suggestions for improvement, or why their approach was the best one for

the goals. (You can’t just say they did a good job. Look critically)

7. Include a photocopy of your selected paper (IT WILL NOT BE RETURNED)

Written papers must be word processed and no more than 2 double-spaced pages. Figures or tables may be included as additional pages, but you can also just refer to them in your narrative since a photocopy of the paper will be included. No handwritten assignments will be accepted.

-------------------

The paper above is written by my teacher

the teacher wants four 4 papers not before 2016 published. each paper have the the 7 points above, not as a paragraph but as a points from one to six while number 7 i will print the full text and attach it with the papers.

Tutor Answer

Proff_White
School: New York University

Attached.

Running head: GENE TRANSFER AND EXPRESSION

Gene transfer and expression

Student’s name:
Institutional affiliation:

1

GENE TRANSFER AND EXPRESSION

2

Gene transfer and expression
1. Overall scientific goal(s)
The overall goal was to find out whether the transfer of recombinant adeno-associated
virus (rAAV) vectors could overexpress the cartilage-specific transcription factor sox9 and the
pleiotropic transformation growth factor beta (TGF-B) to aid in healing and strengthening of the
cartilages if they are damaged. This is because cartilages are very delicate organs in the
movement systems and once damaged, they take time to heal or not heal at all, which affects
moving parts especially body joints.
2. Gene being transferred, how and why
The gene being transferred is the lacZ gene for E. coli β-galactosidase. The gene, which
is found in the recombinant adeno-associated virus (rAAV) is trnsduced into the aspitates taken
from the patient, and then induced back into the patient using the StemPro Osteogenesis
Differentiation kit or the StemPro Adipogenesis Differentiation kit. The genes are transferred
into the patient to help in initiating proliferation, chondrogenic differentiation and matric
synthesis. It is actualy transferred to help in th healing process of the cartilages when they are
damaged.
3. Vector specifics (such as plasmid, regulatory cassettes, etc)
The vector used is mainly plasmids and the rAAV vector. The rAAV was packaged as a
conventional vector. It mainly used the 293 adenovirus transformed embryonic kidney cell line.
The adenovirus 5 was used to provide helper functions, together with the pAd8 helper plasmid.
They combined to provide safe transfer and action of the transferred genes.
4. Recipient specifics (what cells and why)

GENE TRANSFER AND EXPRESSION

3

The specific cells that were targetted were the chondroregenerative cells. These cells are
responsible for the regeneration of body cells and tissues when they are damaged. Since these
cells are adversely ruined during joint or cartilege damage, they usualy take long to heal or never
heal at all. The cells are thus infused with the lacZ gene for E. coli β-galactosidase to help in
improving the regeneration process.
5. Detection of transfer/expression
The detection of the expression of the gene activity is assesed using the β-gal+ cells. The
percentage change of the cells expressing the β-gal+ cells before and fter the gene tranfer would
indicate whether the tranfer was successful or not. Increased cells with the % β-gal+ cells
indicates successful tranfer. The density of the stain in the H&E sections also indicates.
6. Your suggestions for improvement, or why their approach was the best one for
the goals.
The approach used by the researcher is actually innovative and a better approach in
handling cartilage problems. This is because the use of human bone marrow-derived
mesenchymal stem cells to help in healing cartilages is no longer an active practice in healthcare,
which necessitated the adoption of a more effective means of therapy. The use of this gene
therapy has been proved to be more superior and effective in rekindling repair processes in
primary human bone marrow aspirates.

GENE TRANSFER AND EXPRESSION

4
References

Tao, K., Rey-Rico, A., Frisch, J., Venkatesan, J. K., Schmitt, G., Madry, H., . . . Cucchiarini, M.
(2017). Effects of combined rAAV-mediated TGF-β and sox9 gene transfer and
overexpression on the metabolic and chondrogenic activities in human bone marrow
aspirates. Journal of Experimental Orthopaedics. Retrieved from https://jeoesska.springeropen.com/articles/10.1186/s40634-017-0077-5


Running head: GENE TRANSFER AND EXPRESSION

Gene transfer and expression

Student’s name:
Institutional affiliation:

1

GENE TRANSFER AND EXPRESSION

2

Gene transfer and expression
1. Overall scientific goal(s)
The specific goal of this research was to find out whether modulation can be employed in
the manufacturing of vectors and genes transfused into patients during gene therapy to reduce the
overall immunogenicity. This is following the many host immune responses, which has remained
the main barrier to effective use of gene therapy for the treatment of various illnesses. Immune
responses is high among both the in vivo and ex vivo gene therapy approaches.
2. Gene being transferred, how and why
The in in-vivo transfer of genes, it was established that the use of antigen presenting cells
(APC) helped to reduce immune responses, both in the targetted cells and tissues as well as if the
effects of the genes spreads to the bloodstream. On the other hand, ex vivo transfer of genes
necessitaed the use of lympho-depleting regimens, which helped to reduce immune responses.
These strategies helped to reduce the agitation or sensitivity of the body systems to initate an
immune response against the transgene or product.
3. Vector specifics (such as plasmid, regulatory cassettes, etc)
The vector that was investigated was the lentiviral vesctors (LV). These are vectors that
have capability to penetrate both dividing and non dividing cells. These are basically retroviruses
with the capability to permiate through the membranes due to their preintegration complex. They
are made of a pseudotyoe, a a surface protein and a transgene expression cassette. The vesicular
stomatitis virus surface glycoprotein (VSV.G) is the major c...

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Review

Anonymous
Excellent job

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