biocytogen.com
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2606:4700:20::681a:b25
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Submitted URL: https://pg6ec7gbb.cc.rs6.net/tn.jsp?f=001ObgE6iBzFl8rrMeF9Qqkq_-4n0WTmOn8vNqAc1OQolxM9NHMAHUrr6U3S5-ZPhCrUr8illh2KNgMTulqGi9q...
Effective URL: https://biocytogen.com/gene-targeting-service-verified-with-industry-leading-quality-control-measures/
Submission: On August 13 via api from FR — Scanned from FR
Effective URL: https://biocytogen.com/gene-targeting-service-verified-with-industry-leading-quality-control-measures/
Submission: On August 13 via api from FR — Scanned from FR
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</form>POST /gene-targeting-service-verified-with-industry-leading-quality-control-measures/#wpcf7-f4880-o1
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<h3>Empowering Health Sciences News</h3>
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<p>Sign up for our monthly newsletter to stay up to date on the latest news.</p>
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</form>Text Content
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* Necessary 19
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page navigation and access to secure areas of the website. The website cannot
function properly without these cookies.
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CookieConsentStores the user's cookie consent state for the current domain
Expiry: 1 yearType: HTTP Cookie
1.gifUsed to count the number of sessions to the website, necessary for
optimizing CMP product delivery.
Expiry: SessionType: Pixel Tracker
* Google
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personalization and measuring advertising effectiveness.
test_cookiePending
Expiry: 1 dayType: HTTP Cookie
_GRECAPTCHAThis cookie is used to distinguish between humans and bots.
This is beneficial for the website, in order to make valid reports on the
use of their website.
Expiry: 180 daysType: HTTP Cookie
_grecaptchaThis cookie is used to distinguish between humans and bots.
This is beneficial for the website, in order to make valid reports on the
use of their website.
Expiry: PersistentType: HTML Local Storage
rc::aThis cookie is used to distinguish between humans and bots. This is
beneficial for the website, in order to make valid reports on the use of
their website.
Expiry: PersistentType: HTML Local Storage
rc::bThis cookie is used to distinguish between humans and bots.
Expiry: SessionType: HTML Local Storage
rc::cThis cookie is used to distinguish between humans and bots.
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rc::d-15#This cookie is used to distinguish between humans and bots.
Expiry: PersistentType: HTML Local Storage
rc::fThis cookie is used to distinguish between humans and bots.
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__cf_bm [x2]This cookie is used to distinguish between humans and bots.
This is beneficial for the website, in order to make valid reports on the
use of their website.
Expiry: 1 dayType: HTTP Cookie
_cfuvid [x2]This cookie is a part of the services provided by Cloudflare -
Including load-balancing, deliverance of website content and serving DNS
connection for website operators.
Expiry: SessionType: HTTP Cookie
* LinkedIn
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bcookieUsed in order to detect spam and improve the website's security.
Expiry: 1 yearType: HTTP Cookie
li_gcStores the user's cookie consent state for the current domain
Expiry: 180 daysType: HTTP Cookie
bscookieThis cookie is used to identify the visitor through an
application. This allows the visitor to login to a website through their
LinkedIn application for example.
Expiry: 1 yearType: HTTP Cookie
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elementorUsed in context with the website's WordPress theme. The cookie
allows the website owner to implement or change the website's content in
real-time.
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wpEmojiSettingsSupportsThis cookie is part of a bundle of cookies which
serve the purpose of content delivery and presentation. The cookies keep
the correct state of font, blog/picture sliders, color themes and other
website settings.
Expiry: SessionType: HTML Local Storage
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way the website behaves or looks, like your preferred language or the region
that you are in.
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lidcRegisters which server-cluster is serving the visitor. This is used in
context with load balancing, in order to optimize user experience.
Expiry: 1 dayType: HTTP Cookie
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tidio_state_# [x2]Remembers whether the user has minimized or closed
chat-box or pop-up messages on the website.
Expiry: PersistentType: HTML Local Storage
* Statistics 10
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with websites by collecting and reporting information anonymously.
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_hjSession_#Collects statistics on the visitor's visits to the website,
such as the number of visits, average time spent on the website and what
pages have been read.
Expiry: 1 dayType: HTTP Cookie
_hjSessionUser_#Collects statistics on the visitor's visits to the
website, such as the number of visits, average time spent on the website
and what pages have been read.
Expiry: 1 yearType: HTTP Cookie
hjActiveViewportIdsThis cookie contains an ID string on the current
session. This contains non-personal information on what subpages the
visitor enters – this information is used to optimize the visitor's
experience.
Expiry: PersistentType: HTML Local Storage
hjViewportIdSaves the user's screen size in order to adjust the size of
images on the website.
Expiry: SessionType: HTML Local Storage
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__hsscIdentifies if the cookie data needs to be updated in the visitor's
browser.
Expiry: 1 dayType: HTTP Cookie
__hssrcUsed to recognise the visitor's browser upon reentry on the
website.
Expiry: SessionType: HTTP Cookie
__hstcSets a unique ID for the session. This allows the website to obtain
data on visitor behaviour for statistical purposes.
Expiry: 180 daysType: HTTP Cookie
hubspotutkSets a unique ID for the session. This allows the website to
obtain data on visitor behaviour for statistical purposes.
Expiry: 180 daysType: HTTP Cookie
* LinkedIn
1
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AnalyticsSyncHistoryUsed in connection with data-synchronization with
third-party analysis service.
Expiry: 30 daysType: HTTP Cookie
* YouTube
1
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yt-player-headers-readableUsed to determine the optimal video quality
based on the visitor's device and network settings.
Expiry: PersistentType: HTML Local Storage
* Marketing 43
Marketing cookies are used to track visitors across websites. The intention
is to display ads that are relevant and engaging for the individual user and
thereby more valuable for publishers and third party advertisers.
* Google
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Some of the data collected by this provider is for the purposes of
personalization and measuring advertising effectiveness.
_gaUsed to send data to Google Analytics about the visitor's device and
behavior. Tracks the visitor across devices and marketing channels.
Expiry: 2 yearsType: HTTP Cookie
_ga_#Used to send data to Google Analytics about the visitor's device and
behavior. Tracks the visitor across devices and marketing channels.
Expiry: 2 yearsType: HTTP Cookie
_gatUsed to send data to Google Analytics about the visitor's device and
behavior. Tracks the visitor across devices and marketing channels.
Expiry: 1 dayType: HTTP Cookie
_gcl_auUsed by Google AdSense for experimenting with advertisement
efficiency across websites using their services.
Expiry: 3 monthsType: HTTP Cookie
_gidUsed to send data to Google Analytics about the visitor's device and
behavior. Tracks the visitor across devices and marketing channels.
Expiry: 1 dayType: HTTP Cookie
pagead/landingCollects data on visitor behaviour from multiple websites,
in order to present more relevant advertisement - This also allows the
website to limit the number of times that they are shown the same
advertisement.
Expiry: SessionType: Pixel Tracker
NIDPending
Expiry: 6 monthsType: HTTP Cookie
collectUsed to send data to Google Analytics about the visitor's device
and behavior. Tracks the visitor across devices and marketing channels.
Expiry: SessionType: Pixel Tracker
#,#Collects data on user behaviour and interaction in order to optimize
the website and make advertisement on the website more relevant.
Expiry: PersistentType: HTML Local Storage
#,#_expiresAtCollects data on user behaviour and interaction in order to
optimize the website and make advertisement on the website more relevant.
Expiry: PersistentType: HTML Local Storage
* Hubspot
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__ptq.gifSends data to the marketing platform Hubspot about the visitor's
device and behaviour. Tracks the visitor across devices and marketing
channels.
Expiry: SessionType: Pixel Tracker
* LinkedIn
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li_sugrCollects data on user behaviour and interaction in order to
optimize the website and make advertisement on the website more relevant.
Expiry: 3 monthsType: HTTP Cookie
UserMatchHistoryEnsures visitor browsing-security by preventing cross-site
request forgery. This cookie is essential for the security of the website
and visitor.
Expiry: 30 daysType: HTTP Cookie
* YouTube
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GENE TARGETING SERVICE VERIFIED WITH INDUSTRY-LEADING QUALITY CONTROL MEASURES
Home News & Resources Posts Gene Targeting Service Verified with
Industry-leading Quality Control Measures
GENE TARGETING SERVICE VERIFIED WITH INDUSTRY-LEADING QUALITY CONTROL MEASURES
At Biocytogen, we take pride in our quality control measures when we target and
edit genes for our mouse models and drug development endeavors. From preventing
potentially disastrous results before they occur to instituting reliable,
tried-and-true protocols, proper attention to quality control when gene-editing
can save clients both time and resources. For example, based on our internal
data, the random insertion rate of gene-edited mice generated by CRISPR based
EGE method is about 32%, and almost half of them can’t be segregated from the
properly targeted allele through breeding.
Quality control, we believe, is integral and vitally important not only in
matters of quality assurance during the model production steps but also in the
end-user application process. The standard procedures involved in the QCs may
seem costly, labor-intensive and time-consuming, but it is absolutely rewarding
from a long-term perspective. To discuss some aspects of quality control in gene
editing, we would like to introduce you to two expertise, in-house scientists at
Biocytogen: Min Deng, Ph.D. and Dan Wang, Ph.D.
WHO ARE YOU?
Min Deng, Ph.D. (MD) was a research assistant professor and manager at the
University of Rochester, specializing in neuroscience, development, transgenic
and knockout technologies. At Biocytogen, Min is a senior scientist II and a
senior project manager. She has more than 18 years of expertise in the
production and application of gene-edited cell lines and rodent (mouse and rat)
models. She is keen on anything related to biology. She likes music, travel and
good food.
Dan Wang, Ph.D. (DW) graduated from Cornell University and obtained postdoctoral
training at Harvard Medical School, specializing in diabetic embryopathy, stem
cell, and regenerative medicine. Upon joining Biocytogen, Dan has been building
up the company’s expertise in genome editing and program management. Dan is a
senior scientist providing targeting strategy designs and a project manager
leveraging her technical expertise, scientific conceptual understanding, and
management capacity to provide strategic support with total solutions to
accommodate the researchers’ request. Dan thrives working in a dynamic
environment by building multicultural teams as well as driving internal
alignment and external collaborations.
CAN YOU DESCRIBE A TIME WHERE YOU EXPERIENCED A TROUBLESOME SITUATION THAT COULD
HAVE BEEN PREVENTED BY BETTER QUALITY CONTROL?
MD: I used to be a manager in the transgenic core at University of Rochester.
When we first attempted to generate a floxed mouse model using CRISPR/Cas9
method, we didn’t test sgRNA activity in vitro. Instead of targeting vectors, we
used DNA oligo as the DNA template. After performing several rounds of
microinjection, even though we screened many gene-edited F0 pups, they all had
indels.
DW: Yes, we have experienced several challenging situations since the first
utilization of our CRISPR/Cas9-based EGE™ platform in 2014. Due to the lack of a
proficient QC system at the time, several models generated in the first batch
were problematic. Undesired phenotypes were observed, which theoretically
wouldn’t be resulted from the designed gene manipulation itself, suggesting that
undesired cuts or random integrations might have occurred. In response, we have
reinforced and optimized the system since then by adding three important QC
steps to correct the situation. We are quite confident that we can now deliver
the models with premium quality for our collaborators.
WHAT ARE THE QUALITY CONTROL SYSTEMS IN PLACE AT BIOCYTOGEN?
Biocytogen offers three important quality control systems when the
CRISPR/Cas9-based EGE™ technology is applied:
1. Southern blot analysis is performed during F1 mice screening to exclude the
possibility of random integration – we only deliver F1 mice with a clean
genetic background without random insertion issues. Our data showed that
when the ES cell method is used for the preparation, the probability of
random insertion is about 20%; if CRISPR/Cas9 is used, the probability of
random insertion increases to 32%. This QC step is vitally important;
2. Two rounds of PCR and sequencing analysis are performed on F0 founder stage
to screen out undesired genome edits; two rounds of PCR and sequencing
analysis are performed on F1 generation to ensure the whole targeted region
are correct, excluding the ones with undesired genome edits and keeping the
ones with desired edits only;
3. Highly predicted off-target site screening is performed for F1 mice. For
each targeting site, we design at least eight sgRNAs and test their activity
in vitro using our UCA kit to determine one appropriate sgRNA with high
specificity and high activity, which dramatically decreases off-target
effects.
WHAT MAKES QUALITY CONTROL AT BIOCYTOGEN STAND OUT?
MD: For our improved CRISPR/Cas9 based EGE system, instead of an oligo DNA
template, we co-inject the targeting vector DNA with Cas9 mRNA and sgRNA. The
targeting vector includes 5’ and 3’ homologous arms (~ 1.5 kb) which reduce
random insertion and off-target effects tremendously.
DW: There are many things that make us stand out. If you want me to pick one, I
would say the Southern blot analysis for F1 generation of mice generated by
CRISPR/Cas9-based EGE™ technology. So far Southern blot analysis is the most
effective way to exclude the F1 mice with random integration issues.
Unfortunately, most other vendors either haven’t realized that or could not
afford to perform Southern blot analysis, which requires a substantial amount of
know-how to master.
Indeed, research has demonstrated that even in on-target mutagenesis cases, DNA
breaks caused by CRISPR/Cas9 may result in large-scale deletions or other
undesired results (Kosicki). Improvement in confirming on-target alleles of
interest is still needed (Ayabe). Utilizing the lessons learned and wisdom
gained from over a thousand completed past projects, our scientists are focused
on providing our clients with the most up-to-date, innovative protocols,
technologies, and strategies.
Figure 1. CRISPR/Cas9, while specific and precise in many cases, can also
produce random insertions, deletions, and inversions. We believe Southern Blot
is the most effective way to exclude these undesired results. Image from:
Biocytogen.
Biocytogen offers gene-editing services based on multiple modalities, including
CRISPR/Cas9 Extreme Genome Editing (EGE™), ESC-based gene editing, and
transgenic technology, all backed with our industry-leading quality control as a
top priority. With a team of over 200 scientists in China in addition to those
in Boston, MA USA, Biocytogen is a global, multi-cultural company that is well
versed in collaboration and communication. You can find Biocytogen’s work in
numerous publications in high profile journals such as Nature, J. of Immunology,
and Oncogene.
Download Gene Editing Services Brochure
Learn More about Target Strategies
Works Cited
Kosicki, M., Tomberg, K., & Bradley, A. (2018). Repair of double-strand breaks
induced by CRISPR–Cas9 leads to large deletions and complex rearrangements.
Nature biotechnology, 36(8), 765-771.
Ayabe, S., Nakashima, K., & Yoshiki, A. (2019). Off-and on-target effects of
genome editing in mouse embryos. Journal of Reproduction and Development, 65(1),
1-5.
Updated March 16 2020.
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