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wellgenetics.com [email protected]+8862-2651-1809 +8862-2782-9911
CRISPR Genome EditingCRISPR Genome EditingEmbryo MicroinjectionEmbryo MicroinjectionService CatalogService Catalog
CRISPR Genome EditingEmbryo MicroinjectionService Catalog
CRISPR Genome Editing Services(Drosophila)
Embryo Microinjection Services(Drosophila & Mosquito)
Free Consulting Service: Strategy & Plan
Below are 4 most popular genome editing requests: Gene Disruption, Reporter Knock-in, Tag Knock-in,
and Seamless Editing. Any special editing request other than listed is welcomed. WellGenetics scientists
will suggest strategies to meet specically to your needs and propose a most practical and economic way
to achieve what you want!
A
dsDNA Donor
// //
loxP loxP
stop Selection Marker
HDR
Genetic Screening
// //stop Selection Marker
B
Cassetteoptions for Ⓐ
B1loxP
stop
loxP
stopattPX
B2
B3
loxP loxP
attP
loxP loxP
attP attP
B4
B5
Reporter
GFP, Gal4,Gal4::VP16,LexA ...
loxP loxP
attP
Reporter
GFP, Gal4,Gal4::VP16,LexA ...
loxP loxP
attP attP
Cassetteoptions for Ⓑ
Gene Disruption
// //
HDR
Genetic Screening
dsDNA Donor
attP attP
// //
loxP loxP
WellGenetics is dedicated to providing researchers professional
services in generating transgenes and gene knockout/knockin in
y models. We are an independent company from any academic
research institutes. Each project will be designed, executed, and
supervised by postdoctoral scientists, thus to ensure the quality
and reliability of entire experiment process.
Simply let us know the name of gene and how you want it to be
edited. We will propose the best strategy and do the rest of steps
for you - from Design to Fly! Please see service lists below.
Fly Genome Editing Expert — WellGenetics
A2loxP
stop
loxP
stopattPX
A3 hsp70 wReporter
GFP, Gal4,Gal4::VP16,LexA ...
loxP loxP
BglII
A1loxP
stop
loxP
stop
T2A
Selection Marker
Selection Marker attPattPhsp70 w
hsp70 w
3xP3 RFP
3xP3 RFP
3xP3 RFP
3xP3 RFP
3xP3 RFP
Selection Marker Options: SM1 3xP3 RFP3xP3-RFP SM2 hsp70 whsp70-w cDNA
Reporter Knock-in
E Gene Manipulations: T-GEM/ CRIMIC (CRISPR + MiMIC) (Diao et al., 2015 Cell Report)
// //coding intron
T-GEM Plug In
attP attPSASA SASDattP
Use Crispr/Cas with T-GEM
attP attPattP
pASAattP attPpASAattP
Use "off-the-shelf" MiMIC line OR
// //attP attPattP
attP attBattB
QF2attP attBattB
LexA::QFAD
attP attBattB
3XGal80
attP attBattB
Gal4DBD
attP attBattB
p65AD
Trojanexon
Cassetteoptions for Ⓔ E1 3xP3Gal4 pASA RFP
loxP loxP
attP attP
D Cassetteoptions for Ⓓ
D1 Reporter
GFP, Gal4,Gal4::VP16,LexA ...
loxP loxP
BglII
C C1
C2
C3
Reporter
GFP, Gal4,Gal4::VP16,LexA ...
loxP loxP
Reporter
loxP loxP
attP
Reporter
loxP
attP attP
Cassetteoptions for Ⓒ
T2A
T2A Gal4
// //
dsDNA Donor
loxP loxP
Reporter
// Reporter //Selection Marker
Selection Marker
HDR
Genetic Screening
// //
// //
dsDNA Donor
loxP loxP
Reporter
Reporter
T2A Selection Marker
Selection MarkerT2A
HDR
Genetic Screening
T2A
loxP
Tag Knock-in
GF
// //
Cre/loxP Excision (Add-on Service)
//
//
//
dsDNA Donor
loxP loxP
Tag
Tag
//Tag
HDR
Genetic Screening
Selection Marker
Selection Marker
HA, APEX2, GFP ...
// //
Cre/loxP Excision (Add-on Service)
//
//
//
dsDNA Donor
loxP loxP
Tag
//
HDR
Genetic Screening
Selection Marker
HA, APEX2, GFP ...
Tag
Tag
hsp70 w
hsp70 w
hsp70 w
hsp70 wN-terminal Reporter
C-terminal Reporter
N-terminal Tagging N-terminal Tagging
Linker peptide after excision
Selection Marker
Tag Knock-in (continue)
J
I
L
//
// //
//
ssODN or dsDNA Donor
Seamless Editing (Point Mutation, Fragment Knock-in/ Knock-out/ Replacement)
HDR
PCR/ HRMA/ Digestion Screening (Add-on Service)
M
//
//
//
PBac Transposition (Add-on Service)
dsDNA Donor
PiggyBac-based Scarless System(Developed by O'Connor-Giles' Lab)
TTAA
3xP3 DsRed TTAA
//
TTAA //
//
TTAA
3xP3 DsRed TTAATTAA
HDR
Genetic Screening
K
// //
dsDNA Donor
Tag
//
HA or Flag
loxP loxP
Tag //
HDR
Genetic Screening
Selection Marker
Selection Marker
// //
Cre/loxP Excision (Add-on Service)
dsDNA Donor
// //Tag SDSA
// //Tag SDSA
loxP loxP
Tag SDSA
HDR
Genetic Screening
Selection Marker
Selection Marker
HA, APEX2, GFP ...
gRNA Transgenic Fly
Generation Service
Microinject gRNA plasmid into
requested landing site and set
up the gRNA transgenic stock.
Gene Synthesis Service
To avoid confusion of polymor-
phism or PCR errors occurred in
PCR cloning, you may choose to
synthesize homology arms.
Selection Marker Excision Service
To excise selection marker oxed by
loxP/PBac, we provide cre/loxP
excision and PBac transposition
services.
Customized Cassette
Cloning Service
Design and construct
cassette for your specic
research needs.
PCR Screening Service
Individual F1 progeny is back-
crossed to balancer. We then
perform single-y genomic
PCR on each fertile F1 ies.
HRMA Screening
Individual F1 progeny is back-crossed to
balancer for high resolution melting analysis
(HRMA). Using HRMA to screen for single-
nucleotide mismatches or indel mutations.
Digestion Screening
Individual F1 progeny is back-crossed to balancer.
We then perform single-y genomic PCR on each
fertile F1 ies. PCR products will be puried and cut
by endonuclease and analyzed by gel eletrophoresis.
Add-on Services for CRISPR Genome Editing
H Internal Tagging C-terminal Tagging
// //
Cre/loxP Excision (Add-on Service)
dsDNA Donor
// //
loxP
Tag
loxP
// //
HDR
Genetic Screening
Selection Marker
HA, APEX2, GFP ...
Tag Selection Marker
Tag
// //
Cre/loxP Excision (Add-on Service)
dsDNA Donor
// //Tag
loxP
Tag
loxP
// //Tag
HDR
Genetic Screening
Selection Marker
Selection Marker
HA, APEX2, GFP ...
C-terminal Tagging C-terminal Tagging
Linker sequence after excision Linker peptide after excision
CRISPR Service Packagewith Selection Marker -or- with PCR Screening
WellGenetics provides CRISPR Service Package including Design, Cloning,
Microinjection, Screening and Stock Establishment to bring you gene-edited
ies. Simply let us know the name of gene and how you want it to be edited. We
propose the best strategy and do the rest of steps for you - from Design to Fly.
Using homology-dependent repair (CRISPR/HDR), we can knockin desired
cassette in combination with cuts by 1 or 2 guide RNAs. We welcome any project
that requires customized designs to t your specic research needs. Feel free to
show us your interests in CRISPR y!
Workow of CRISPR Service Package
CRISPR Service Package with Selection Marker CRISPR Service Package with PCR Screening
Step 2
Step 1
Select guide RNA target sites at your preferred region(s)
Propose and quote you the best suggestions to your needs
Step 3
Step 4
Design repair template as donor carrying chosen knock-in cassette
Conrm target sites by genomic PCR in the injection strain
Step 5 Guide RNA plasmid construction and plasmid donor construction or ssODN donor synthesis
Step 6 Plasmids DNA preparation & purication
Step 7 200-embryos microinjection
Step 8 G0 cross
Step 9 F1 Screening by visible marker(NO additional charge)
F1 Screening by PCR(with additional charge)
Step 10 Genomic PCR validation
Step 11 Positive line isogenization and balancing
Step 12 If the rst trial fails, free second round microinjection with optimization
If both trials fail, we only charge half of the package price* *Terms and Conditions apply
Package
CRISPR Service PackageYou Provide: Name of Gene & Editing Ideas. We Do the Rest: CRISPR Strategy Proposal, Target Site Design,
Target Site Conrmation, Cloning, Microinjection, Screening , Molecular Validation, and Balancing.
1 week 2+4 weeks 2 weeks 4-5 weeks 4-5 weeks
Step 2-3. Design Step 4-5. Cloning Step 6-7. Microinjection Step 8-9. Screening Step 10. Validation Step 11. Balancing
Our Clients... It is our great honor to make contributions to your research projects
sni t i t u a l r o s et
B i o l o g i e
National Chung Hsing University
Guarantee
// //
Drosophila & Mosquito Embryo Microinjection ServicesWellGenetics provides Fly Microinjection Services for CRISPR, PhiC31, P-element and
PiggyBac transgenesis in Drosophila melanogaster, and Mosquito Microinjection Services
for CRISPR and Transposon transgenesis in Aedes aegypti.
Embryo Microinjection
Microinjection Service Lists
PhiC31 Microinjection & Screening Services*
200-embryos microinjection, G0 cross & F1 screen (w+).
We return unbalanced positive lines.
We have VK33, VK37, attP2, attP40, ZH51C &
ZH86Fb as regular panel of landing sites. You can
also choose any other sites listed in Bloomington Stock
Center and allow two generations of amplication time.
We guarantee 1-3 transformants for plasmid at standard
size (<13kb).
Bac (P[acman]) Microinjection & Screening
Services*
600-embryos microinjection, G0 cross & F1 screen (w+).
We return unbalanced positive lines.
CRISPR Microinjection
200-embryos microinjection. We return survived larvae
after microinjection.
Mosquito Microinjection
500-embryos microinjection. We return injected embryos
after microinjection.
Non-melanogaster Microinjection
200-embryos microinjection. We return survived pupae
after microinjection.
P-element Microinjection & Screening Services*
200-embryos microinjection, G0 cross & F1 screen (w+).
We return unbalanced positive lines.
We guarantee 5-8 transformants for plasmid at standard
size (<13kb).
*NO guarantee for large plasmid (>13kb).
Add-on Services
Balancing Service
Isogenize & balance positive lines from screening.
We establish y stocks for you.
Plasmid Construction Service (<3kb)
Maximum 3 PCR-fragments and together length below 3kb.
We use High-Fidelity Phusion DNA Polymerase to amplify
the fragments and sequence through all parts we cloned in.
We return bacteria streaking plate and 4µg plasmid DNA.
BAC DNA Prep.
Qiagen Large Construct Kit for BAC DNA preperation
(>20kb) to provide high quality DNA for microinjection.
Validation service by restriction enzyme digestion or PCR
is included!
Plasmid DNA Prep.
Plasmid preperation to provide high quality DNA for
microinjection. Validation service by restriction enzyme
digestion or PCR is included!
Want to Know More about Our Services?Contact WellGenetics [email protected]+8862-2651-1809