T Cell Gene Therapy : The Center Of Research With Stem Cell Transfection In Geney Cells

magine, 20 years from now, sitting in a cold doctor's office deciding the genes of your unborn baby, what color hair, eyes, speed of metabolism, height would you even know what to pick? Impossible you might say but in this day and age technology is growing ever so rapidly that picking the genetic makeup of your baby is closer than you might think. The technology is called CRISPR. This technology doesn't only have the ability to change physical traits, but genetic traits specifically genetic abnormalities and diseases. 20 years ago, no one would have ever thought we would have the answer to, in theory, cure every genetic disease from sickle cell anemia to cystic fibrosis. However, with great scientific breakthroughs comes questioning and

Once the complex was bound to the DNA, a cut would be made to eliminate and destroy the invaders. 83% of archaeal genomes and 45% of bacterial genomes (Shabbir, M. et al, 2016) were shown to be able to successfully utilize the CRISPR Cas9 system. These are very promising statistics, so it is no wonder that there has been such an advancement in the past few years to bring this technology to eukaryotic cells, mammalian cells and eventually human cells.

CRISPR has been garnishing a lot of media attention recently and it is not just popular among the scientific community but also the general public. Several online news outlets and scientific journals have been talking about the significance CRISPR-Cas could have for the field of genetics and science as a whole. I even came across a Youtube video from The Verge, a tech channel that normally does reviews on new smartphones and laptops talking about CRISPR [15]. So why is CRISPR gaining so much attention both from the scientific community and the general public? The answer lies in the potential this technology possesses.

For many years biomedical researchers like myself have been trying to create more proactive ways to amend the genome for living cells. In more recent fieldwork studies there has been a new state of the art instrument based on bacterial CRISP in close works with protein 9 often referred to as CAS9 from the streptococcus progenies have possibly unlocked new data. The CRISP/CAS9 tries to manipulate the function of the gene using homologous recombination and RNA interference, but is set back because it can only provide short term restriction of the genes function and it’s iffy off- target effects.

More recently, Kang et al have employed a different approach using a non-viral delivery method for CRISPR-Cas, known as the

The ability to engineer biological systems and organisms has an enormous potential for applications across basic science, medicine and biotechnology. Genome editing is a group of technologies that allow scientists the ability to change an organism’s DNA, which can provide better outcomes for health and disease control compared to natural immunity and mutations. Genome editing (gene editing) allows genetic material to be added, removed or altered at particular locations in the genome. A number of gene editing technologies have emerged in recent years with one of the most versatile and precise methods of genetic manipulation being Crispr-Cas9 (Steve scott,2016). The term Crispr-cas9, (clustered regular interspaced short palindromic repeats)

CRISPR-Cas9 is a unique technology that can be used to edit the human genome (1). Compared to other techniques of editing DNA, CRISPR and Cas9 are cheaper, faster, and more accurate. CRISPR-Cas9 gives geneticists as well as medical researchers the ability to edit specific parts of the genome by the removal, adding, or altering of certain sections of DNA sequences (1). CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats and is a part of bacterial defense system (2). The two key molecules of CRISPR-Cas9 are Cas9 (an enzyme that can cut DNA at specific locations) and guide RNA (a small piece of RNA sequence that guides where the Cas9 should cut) (1). CRISPR is a crucial component

CRISPR cas9 should continue to advance in the science world because “UC Berkeley researchers have made a major improvement in CRISPR-Cas9 technology that achieves an unprecedented success rate of 60 percent when replacing a short stretch of DNA with another”(Antonio Carusillo, PhD Candidate in Genetic Engineering (Marie Curie) at University of Freiburg (2018-present). This statistic shows that there is more of a chance to success but there is a chance to fail 40 percent but overall it will succeed which is why people are lenient about will it actually work or not, but as technology get better so will treatments to cure hard to pinpoint disease such as cancer, zika, or leukemia. Genome editing (also called gene editing) is a group of technologies that give scientists the ability to change an organism's DNA. These technologies allow genetic material to be added, removed, or altered at particular locations in the genome. Several approaches to genome editing have been developed. A recent one is known as CRISPR-Cas9, which is short for clustered regularly interspaced short palindromic repeats and CRISPR-associated protein. The CRISPR-Cas9 system has generated a lot of excitement in the scientific community because it is faster, cheaper, more accurate, and more efficient than other existing genome editing methods (yin, steph.What Is CRISPR/Cas9 and Why Is It Suddenly Everywhere? published april 30,

We review the history of CRISPR biology from its initial discovery through the elucidation of the CRISPR-Cas9 enzyme mechanism, which has set the stage for remarkable developments using this technology to modify, regulate, or mark genomic loci in a wide variety of cells and organisms from all three domains of life. These results highlight a new era in which genomic manipulation is no longer a bottleneck to experiments, paving the way toward fundamental discoveries in biology, with applications in all branches of biotechnology, as well as strategies for human therapeutics

Gene Editing is a method that can be used to change or edit the genetic code. Researchers and doctors can add or delete a sequence in the genetic code. Adding or deleting a sequence in the genetic code is just like copying and pasting in a word document. Gene editing is being used to cure disease and/or disorders, and has been around for years but has never been very accurate or precise. Now, with the breakthrough technology of the year, the CRISPR-Cas9 gene editing could be easier than ever.The CRISPR-Cas9 has the potential to cure diseases that many have died from. The CRISPR-Cas9 is like a little pair of scissors that will cut out, repair , or even replace DNA. The changes that are made with this machine will be passed on from generation to generation. Like said before, gene editing is not new to the world, but the CRISPR-Cas9 is the first technology being used to undergo gene editing. Before the CRISPR-Cas9 there was the TALENs. Out of all the technology for gene editing CRISPR-Cas9 has been the most precise of them all. As soon as it is perfected it will be the next

By using the CRISPR/Cas9, it is possible to alter DNA to correct genetic diseases in animals and modify DNA sequences in embryonic stem cells (Rogers, 2016). This opened up the process of genome modifications in humans through the germ line (sperm and egg) (Rogers, 2016). Doudna became interested in the CRISPR and found out it is part of the bacterial immune system. It all started with RNA sequences from invading viruses that become part of the bacterial genomes. From there the viral sequences stay as DNA in the spacers between the short repeating blocks of bacterial DNA sequences (Rogers, 2016). When the virus attacks the bacterial cell again, the spacer DNA is converted to RNA. A Cas9 enzyme and a second RNA molecule attach themselves to the new RNA and then searches for a matching strand of viral DNA (Rogers, 2016). The Cas9 cuts the viral DNA which prevents the virus’s replication. Charpentier and Doudna found that the guide RNA sequence could change the direction of the Cas9 to a precise DNA sequence. By this finding, it transformed the genome engineering and could be treatments for human diseases (Rogers,

Surrounded by patent and ethical issues lies a gene editing method with massive potential within the biotechnology industry. The CRISPR-Cas9 system works like ‘molecular scissors’ where Cas9 is an endonuclease that targets a specific DNA sequence (Griggs. 2015). This is more efficient than the previous methods such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), as well as being simpler to use. CRISPR-Cas9 uses single guided RNA (sgRNA) to reach the desired gene, where it is able to cleave the double stranded DNA in the presence of a Protospacer Adjacent Motif sequence (PAM sequence) (Ran et al. 2013). This is the stage where the gene can be altered via the cells own repair system due to the break in the DNA (Nehme et al. 2014). Solutions to sickle-cell anaemia, malaria and beta thalassemia are just a few of the life changing impacts this method could have in the future.

Both “Malaria and HIV-1 are 2 of the most common infections in sub-Saharan Africa and, to a lesser extent, in other developing countries. It is estimated that 38 million Africans are infected with HIV-1, whereas 300 million to 500 million suffer from malaria each year” (Whitworth 3). These two disease go hand in hand to cause the millions of deaths that occur in Sub-Saharan Africa every year. HIV, a disease that weakens immune system by attacking and killing T-cells in the body, allows for Malaria to take a greater and deadlier effect on the already weakened human body. HIV/AIDS might eventually be a thing of the past because CRISPR has been used to get rid of diseases in animals, such as cystic fibrosis, muscular dystrophy and a form of hepatitis and scientists are now using these methods to try to cure AIDS (Michael 2). Curing of HIV or AIDS is no simple task since the disease is extremely resilient, plentiful and is present in the majority of the body’s blood stream. Scientists would have to somehow alter the genes of the body's T-cells to resist being taken over by the HIV or alter the genes of the HIV to not be able to spread and infect other body cells. Genetic engineering is the only method that provides this realistic possibility of being able to treat and cure HIV and AIDS that otherwise would

Hsu, P., Lander, E., & Zhang, F. (2014). Development and Applications of CRISPR-Cas9 for Genome Engineering. Cell, 157(6), 1262-1278. http://dx.doi.org/10.1016/j.cell.2014.05.010

There are three techniques that researchers are working on. The first and most common is ex vivo ( or "outside the living body") therapy. The defective cells are removed from the patient and replaced with the normal DNA before returning to the body. This therapy targets the blood cells because many genetic defects alter the functioning of one type of these cells or another. But since blood cells have limited life spans follow-up treatments are required. Future efforts will most likely target stem cells of the bone marrow. Stem cells are ideal for gene therapy because they appear to be immortal. Researchers have obtained stem cells from human bone marrow, but they are having difficulties getting genes into the cell as well as inducing the cells to produce many new blood cells (Anderson, 1995).