This review illustrates a
comparison of currently implemented strategies and the need of a novel
technique to cure the COVID-19. Current strategies served a lot to cope up with
the situation by decreasing the rate of fatalities i.e. some FDA approved drugs
and giving immunity by the plasma proteins of cured patients, however these
approaches could not eliminate the pandemic from the society emphasizing that
some advance technology need to be considered in order to fight with SARS-COV-2
virus. CRISPR-Cas based approach to inactivate the virus before its fusion with
host DNA has been successful in in-vitro studies performed at Stanford
University along with SHERLOCK & MAMMOTH biosciences who were able to
degrade 80 % viral genome. Here we have also done a literature review of major
affected countries China, Italy, United states, and India.
CRISPR-Cas, Cure, Covid-19, FDA,
Ever since the man first lay his feet
in this world the process of natural selection is going on to keep check and
balance between earth resources and the man utilizing them side by side
sometimes with natural disasters like floods, volcano eruptions ,earthquakes
etc. and sometimes with Epidemic and Pandemic diseases like Cholera ,Influenza
,Typhus, small pox, Measles, leprosy, Polio ,Malaria, HIV and now COVID 19. The
2019 novel coronavirus (2019-nCoV) also known as the severe acute respiratory
syndrome corona virus 2 (SARS-CoV-2) is rapidly spreading from its origin in
Wuhan City of Hubei Province of China to the rest of the world and affecting a
significant world’s population. The infection has spread in more than 200
countries and territories . Till 16/10/2020 around 39,418,121 cases of
coronavirus disease 2019 (COVID-19) and 1,106,128 deaths have been reported .
This review article gives a bird’s eye view about the effect of virus on
different age groups and gender as well as a comparison of conventional and
advanced treatment strategies.Fortunately, children have been least
affected by COVID 19, but the future course of this virus is unknown. Data from
China, United States, Italy, and India was collected and presented in this
review article in tabular or graphical form.
Strategies to Combat
combat COVID-19 numerous strategies are under consideration as well as some are
already applied. i.e. around 300 different ways to cope up with this disease
and more than 200 vaccines are under clinical trials at various stages,
However, traditional approaches look like lagging behind the age to fight the SARS-COV-2
even the antiviral therapy. On the contrary Monoclonal antibodies and
Vaccination are promising but still many hurdles needed to be passed on. An
accurate diagnostic and treatment approach against COVID-19 is far above the
current approaches e.g. clinical trials of vaccines on large scale to develop
safety & efficacy profile and financial burden on R & D of Antivirals
and industrial production hinders the path to eradicate COVID-19.
gene editing technology to visualize and then target the genome of corona virus
can be a promising lead in this scenario.
Advanced Technique of CRISPR-CAS Based Strategy for Corona
regularly interspaced short palindromic repeats (CRISPR) and proteins
associated with it together constitute a family of sequences of DNA present in
certain prokaryotic Organisms in their genomes i.e. bacteria and archaea.
Immune system of such pathogens is characterized with these proteins and helps
to provide protection against RNA from phages, DNA, plasmids, or other DNA
elements that invade their body. Viral genetic material gets destroyed upon
binding with a specific target sequence that is present on specific crRNA
produced in response to the invasion of foreign substance to the body. Clinical
virology has entered a new era by the discovery of this technique to eliminate
the RNA and DNA based viruses and even detection and identification of specific
sequence of nucleotides is already under use.
SHERLOCK biosciences were able to
detect the S gene and Orf1ab while MAMMOTH Biosciences did target N and
E viral genes facilitating the visual readout of the Corona virus genome .
After that many techniques have proven successful to diagnose SARS-COV-2 even
in < 40 minutes by the CRISPR based lateral flow assay. CRISPR/Cas-NER assay
demonstrated 100 percent agreement with PCR in diagnosis of COVID-19 with the
success probability of p <0.001.
In a recent study
, researchers were able to degrade 80-90% of the genetic material of
SARS-CoV-2 using a pool of Cas13d (RNA Targeting CRISPR from Class II) and crRNA
by targeting multiple regions on the genome of coronavirus. This Technology was
coined the name PAC-MAN (Prophylactic Antiviral CRISPR in
huMAN cells). It led to the fact that a pool of 6 crRNA could target all
the strains of Coronaviruses.
Selective and Efficient Delivery of CRISPR-Cas13 in Antiviral
main obstacle in the application of this technique is the delivery of the
components of CRISPR to the viral genome. Current method to deliver the CRISPR
is non specific means target the both infected as well as normal cells that can
cause harmful side effects even in the absence of Viral RNA. Thus there must be
some methods that can deliver strategically CRISPR components in to the
infected cells only before the testing of this techniques on human level.
One way is to
synthesize Cas13 nuclease in such way to have affinity with the viral
particles, hence it will deliver the CRISPR components specifically and
efficiently into the host cell upon the entrance of the virus itself. In this
way the delivery of the virus and the treatment will follow the same path.
CRISPR-cas complex will shred the RNA of the virus when it will be released
into the host cell cytoplasm by the virus, so the Viral RNA will be cleaved
before its translation and replication inside the host cell nucleus. Hence the
virus itself will be the main key to kill itself selectively and efficiently.
COVID-19 by Area, Age and Gender
The COVID 19 infection which is
affecting a significant world’s population is an emerging global health
concern. The infection has spread in more than 200 countries and territories
affecting more than 12 million people till now. Data from some of the countries
were collected and presented in this article in tabular or graphical form.
the city of China was the place where the virus started spreading. It was
identified in December 2019 in Wuhan however the first case traced back to
November 17, 2019.
Graphical Representation of Number of Cases and Deaths within
Different Age Groups in China
According to WHO COVID 19 dashboard
data, there was an almost linear relationship between total COVID 19 cases and
the number of new reported from April till July. The initial spread was very
fast making the number in December from a few hundred to the end of January at
almost 12,000 and then at the end of February, there were 79,000 cases after
that the spread is limited. There were 84,784 COVID 19 cases in China until 25
A total of 72,314 individuals were
diagnosed with COVID-19 as of February 11, 2020. 44,672 cases (61.8%) were the
confirmed cases, 16,186 cases (22.4%) were suspected, 889 cases (1.2%) were
asymptomatic, and 10,567 cases (14.6%) were clinically diagnosed(Zhi, 2020). Most numbers
of patients were 30 to 79 years of age (87%), 10 to 19 years of age were (1%),
and 3% were age 80 years or older. Most of the cases were diagnosed in Hubei
(75%) and most Wuhan related exposures (86%) . Among 44,672 confirmed cases
in China, 22,981 (51.4%) were the males and 21,691 (48.6%) were the females.
The male to female ratio was 1.06:1 in China overall while in Wuhan it was
Figure 2: Comparison
of Percentage of COVID Cases and Deaths among Males and Females of China
A study was conducted which was published in JAMA (The Journal of
American Medical Association) on April 15, 2020, in which they had studied that
almost more than 50% cases were from the Chicago and in those 50% the
percentage of the deaths accounted for more than 70% of black people, although
blacks were almost 30 percent of the total population.
In Louisiana, 70.5 percent of deaths accounted for black
people who represent 32.2 percent of the state population
The Johns Hopkins University and American Community Survey
indicated that out of 131 predominantly black people regions in the USA the
infection rate was 137.5 out of 100,000 people simply 0.1375% and the death
rate was 6.3 out of 100,000 people which was 0.0063%.
The World Health Organization
declared the COVID-19 outbreak a pandemic on March 11, 2020. Among the
1,761,503 cases reported to CDC from January 22–May 30, individual case reports
for 1,406,098 were submitted to CDC case surveillance.
1.Percentage of Covid-19 Cases within Different Age Groups
Age Group (yrs.)
exclusions, data for 1,320,488 (94%) cases were analyzed. The following table
shows the number of patients affected by COVID 19 with different age groups .
The spread of COVID 19 in the United
States was slightly greater in females as compared to males. Among 1,320,488 of
total analyzed cases, 646,358 of them were the males and 674,130 were the
females. But the number of deaths in males was greater than in females. 38,773
out of 646,358 were the reported deaths in males and that is about (6%) while
on the other hand 32,343 out of 674,130 were the reported deaths in females and
that was (4.8%) .
3: Graphical Representation of no of Cases within Different
Age Groups in United States
2.Number of Cases and Deaths among Males and Females in United
No. of Cases
being the part of Europe was also not safe from the terrorizing effects of
COVID 19, till February 2020 there were only 3 cases of COVID 19 and all of
those which were traveled to china recently on February 2020 a person with
aggravated pneumonia due to COVID 19 was diagnosed in northern Italy Lombardy
region who was in his 30s and had no history of abroad within 2 weeks many
other cases were reported then it was hypothesized the virus was in population
since January 2020 but became active in February 2020 .
4: Graphical Representation of Number of Cases Among Various
Age Groups in Italy
to increasing number of cases the Italian Ministry of Health issued some
policies, according to these policies the patients who have more clinical
symptoms have undergone the tests of COVID 19 and people who have mild symptoms
and asymptomatic were left despite accounting for a limited number of tests,
there was a high portion of positive results about 19.3% (positive cases 21,157
of 109,170 tested as of March 14, 2020) also the case fatality rate changed
from 3.1 % on February 24, 2020, to 7.2 % on March 17, 2020, because the mild
symptoms cases and asymptomatic cases also add their share in fatality
percentage after that almost all patients have shown symptoms whether mild or
severe undergone testing .
overall case fatality rate in Italy (7.2%) was three times more than China
(2.3%) and that increasing number was from the people in Italy who were more
than 70 years whereas in China the death rate in people above 70 years of age
was not as much as in Italy and the possible reason can be genetic makeup and
environmental conditions. Chinese people eat more natural foods thus have
a strong immune system which can bypass the virus attack .
to the data collected there were 22,512 active cases in Italy till March 15,
2020 , the number increased to 105,792 on April 1, 2020, (2) and according
to WHO COVID 19 dashboard data there were 238,159 cases till June 20, 2020 and
the numbers were 245,590 cases until July 25, 2020.
According to Istituto
Superiore di Sanità Italy has 235,579 confirmed cases of COVID-19 and 32,722
associated deaths as of June 8, 2020. The median age of cases is 61 years, and
more than two-thirds of these patients had diabetes, cardiovascular diseases.
Of the patients who died, 38.4% were aged >70 years, 30.9% were aged 51 to
70 years, 28.5% were aged 19-50 years, and 2·2% were aged 0-18 years .
Table 3. Percentage
of Deaths and CFR Among Various Age Groups in Italy
No. of Deaths
% of Deaths
The spread of coronavirus in Italy was
slightly greater in females as compared to males. As the chart shows, 54.2%
were the females and 46.8% were the males. As of May 26, 2020, a study on
32,000 patients revealed that the fatality rate is much higher for men. If the
mortality rate for female patients was 10.4% the mortality rate for male
patients was 17.7% .
first case reported in India was on Jan 30, 2020. As of June 16, 2020, India
had a total of 344,407 confirmed cases of coronavirus and 9,921 deaths(Coronavirus
| 42% of patients in India between 21 and 40 years, 2020).The
Union Health Ministry on April 4, 2020, throwing light for the first time on
the demography of the cases in India, and according to that, about 42% of
confirmed COVID-19 in India ware between the ages of 21-40 years old. The
percentage of those who were over age 60 was only 17%. The U.S. Centers for
Disease Control said that 29% of the affected were the age of 20-44 years and
32% were above 60 years. The United States recorded the 279,355 cases and 7,451
deaths compared to the 2,904 cases and 75 deaths as on April 4, 2020 .
Today's Data intelligence Unit on 3rd April 2020 showed 1801 COVID-19 confirmed
cases revealed that around 60 percent of the patients were below 50, so young
Indians were at higher risk to get this disease as compared to other
population. Similarly, data also revealed that 17% of the patients were in
their 40s, 21% (376 cases) were in their 20s and 22% (391 cases) were in
between 30 and 39 years of age. .
Early reports from countries, such as
China and Italy, have shown that the most vulnerable category affected by
COVID-19 is that of older people. But in India, patients over the age of 60
were only 19% of the total confirmed cases while people aged 80 and older
account for less than 2% of the total confirmed cases of COVID-19 . The CFR (Case Fatality rate) among males was 2.9%,
while that for females it was 3.3% in India. As of May 20, 2020, 66% were the
males affected by COVID-19 infection while 34% were the females.
Conventional Strategies to Treat COVID-19
Representation of %age of cases in different age groups in
study has some limitations as only those countries were included in the study
that were most affected while those countries who had minor impact were
excluded from the study. Hence the results interpreted including the variation
of cases among different age groups and among gender are based on four
countries China, Italy, United States and India.
Secondly the application of CRISPR-Cas
although looks promising but still itself is a barrier to be implemented on
large scale as In-vivo studies and Clinical and Pre-Clinical trials will cost
much time and finance.
future trajectory of this virus is still not clear. From the data, it can be
concluded that most of the patients were 30 to 70 years of age. Age is one of
the important factors playing a key role in the progression of COVID 19.
Different countries have different ratios of young to old age persons. The
recovery rate is also higher in countries with a high number of young people as
compared to those with a high number of old people. The least affected age
group in the world is 0 to 20 years of age. On the other hand, cases in males
in China, Italy, and India are more as compared to females while in the United
States, females are more affected by COVID 19. Different countries and
organizations of the world are trying to discover the treatment of COVID-19 but
nobody has succeeded to discover the effective drug or vaccine against this
rapidly spreading virus. One of those approaches is targeting the genome of
coronavirus by applying CRISPR-Cas associated gene-editing technology.
In the scientific community CRISPR can
be a very crucial and important area of research study to explore the disease
diagnosis as well as treatment in the future with promising potential. Using
this technique, we can develop such Novel drugs to cure a range of diseases for
example Alzheimer's, HIV/AIDS, Cancer, HIV and such pandemics like Covid-19.
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