ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet 1 of 13
`
`MRO REFS 4(768bp)
`
`MIRBOALBFS 1485p
`
`
`
`MIR6O4 WELBFS 4IRBES 1
`1582 bp
`
`FIG. 1
`
`
`
`Replacement Sheet
`
`Sheet 1 of 13
`
`MRO REFS 4(768bp)
`
`MIRBOALBFS 1485p
`
`
`
`MIR6O4 WELBFS 4IRBES 1
`1582 bp
`
`FIG. 1
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet 2 of 13
`
`Donor DNA
`
`
`
`Homology Arn
`
`
`_ Cecaos |_ pityNos
`
`
`
`
`Safe harborlocus#4 faReo4 evevent insertion site}
`
`Qf Site-directed nuclease (e.g. TALEN
`J\ and CRISPR-Cas9}
`
` pr
`HomologyArm| prOMPeCrySANos
`|_itchNos
`| prUbit-oPillos
`{pgmLACEINETSESINOEAe
`a wnprabe (rCMP)
`Probe {cPsae
`ecm i
`
`
`A
`ANINT Probe (native sequences
`Bsu36i
`flanking homology ars }
`
`
`A
`Bsu3Gl
`
`\
`
`Targeted insertion event in the safe harbor locus #1
`
`FG. 2
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet 3 of 13
`
`
`
`FG.3
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet 4 of 13
`
`
`
`oOAIQWICaG{}JOdsanig#GAY
`
` axeontmM——_—
`
`FIG. 4
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet 5 of 13
`
`12345678
`
`
`
`Probe 1
`
`ase
`
`veut
`MKD
`
`
`
`
`
`12343678
`
`12345678
`
`FG. 5
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet6 of 13
`
`She-ditactad tuclaase
`dleavage al the Mlarget
`
`
`
`eachallele
`
`{
`
`0
`
`0
`
`Small inDel in one
`aliale
`
`Small InDelinboth
`alleles,
`
`Small InDel and
`large deletion in
`
`FIG. 6
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet 7 of 13
`
`Detecting targeted mutagenesis: Taqman assay design
`
`
`
`
`ea mnemympainn
`spree
`.
`eeNUedlagste~ nameenananes
`
` ANY ANY
`CTOONGR CACTCST TMAGRAGGTN ASSAGTAGTCAGGHG AGROTIGITINGA CMGRTNTS
`GUGACOCETT GTEGGAQCA BCGSTTTOR CGIRGGGACG THACGTAACG TUACGIOCTS TOCTGRanh ANCAAATCOT GOCTACGAC
`aviare
`
` ERE RT CO ERR*5%‘
`
`:
`
`‘‘%‘‘%
`$
`SAS‘‘;
`:
`:
`‘
`:
`gag tt me wea
`eS Qe TE
`:
`eed
`:
`ot
`i
`fh
`f
`y.
`‘
`i
`:
`.
`Baarauinniinnniiiiniiiiiiiiniiiinittiint iiniitttitinnnintiie eitetttbnntitinttitnttithettUE GSUULERASURERUNGHENNSCUSAR GuaGEAR
`:::
`‘..
`:
`:
`‘
`gt
`oe
`:
`:
`7
`‘
`‘
`:
`2.
`.
`*
`:
`ae:
`5
`‘
`eh.
`SASKANAGAANRGAESAEAAASASCAAGANEE HSEGACSHEGEASGERGAARECGEGHEE
`
`:
`NENW}NNERANNESSNNER
`
`NSSNNNRERN
`
`SNSS
`
`;
`
`’
`
`ANNAN
`
`SSS
`
`ANNREAD
`
`rat
`
`ag
`
`sEAN
`
`FG.7
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet 8 of 13
`
`(A) Target sequence status in different types of events
`
`I, Event without targeted insertion
`
`i], Event with targeted insertion
`
`t
`
` a0)
`
`al
`
`
`
`cn
`AC
`
`
`
`
`~
`
`seesseecenseneenseney,
`
`Lindel | Large deletion
`
`EOI
`
`largeted insertion
`
`FIG. 8A
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet 9 of 13
`
`(B) Copy numbercall of different types of events
`
`
`Event with no targetedinsertion
`
`a} WT no mutation in both allies
`
`b} Small inde! n one allele
`
`©} Sinall indelin both alleles
`
`df} Small indel in oneallele
`and largedeletion in anotherallele
`¢} Large insertion in both alleles
`
`Event with targeted insertion
`
`i} No mutation i one aliais, targeted
`insertion in another allela
`
`g} Small indel in each alieie, targeted
`msenion in anotherallele
`
`h} Large deletion in in each allele,
`targeted insertion in another allele
`
`{i Targeted insertion in both alleles
`
`é
`
`2
`
`2
`
`{
`
`Q
`
`‘
`
`,
`
`Q
`
`oe
`
`FIG.8B (cont'd.)
`
`2
`
`{
`
`0
`
`(
`
`}
`
`j
`
`}
`
`Q
`
`ma
`
`>
`
`>d
`
`>d
`
`>)
`
`>
`
`>4
`
`of
`
`if
`
`$3
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet 10 of 13
`
`NOS (253 bp)
`|
`piZmlbit (1998 bp)
`|
`cPM-O1 (1176 bp)
`(
`INOS (253 bp)
`LBFS (1786 bp)
`
`©
`
`7
`
`
`prZmNfTL (2556 bp)
`RBFS(1451 bp}
`
`\
`
`menGA (1797 bp)
`
`|
`|
`
`
`
`MIR604 RBFS_pNOV2130 T-DNA insert_LBFS
`{1622 bp
`
`FG. 9
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet 11 of 13
`
`TALEN cul
`yi
`<
`
` MIR604 transgenic locus
`em(nsa | PM
`
`sesaarieeessdonniePivaGeUs WY
`
`y11y11
`
`
`\
`
`\
`\
`
`
`oveenesennensennmnogeniny enanennrnnanennnnanennannanensen MARANA
`EBES
`oa
`Bovwnnnnneny
`fevercecee.
`Benen.why
`_hasnnennnnnnnnnnnnasinnnniNSIS NILe.me
`| orbit|cP
`~
`IC Exp. Cass,
`| EPSPS Exp, cass.
`| MI
`Sorceteranatsts
`Donor DNA
`4
`\
`(22872)
`\
`\
`‘\wassvunnsunnnnennans,aassennnsonnnnnnnnne?
`
`SOSAKneAKRAM
`
`sarahiniNNNRNAROANA
`‘
`:
`;
`‘
`Lassasanaunnninn!
`
`\
`
`LS bp
`
`Targeted integration
`
`utUra
`
`
`
`
`
`12 Kb
`
`«i
`
`PA (FE35034)
`P2 {FE4793}
`&NY
`i4444‘MS
`
`prUbil|cP) ICEyp.Cass / EPSPS Ep. Cass, |
`| mCry T7
`MTL
`peewerereceBeeccceece
`fom
`dmnnsey,
`%
`:
`ny
`:
`&
`‘
`
`
`oofcommnanmemneaneinemenancnnnemenanenteatReenTwlehomonnrconndOMRON AAA
`soon
`ay
`PL{FEATIG |
`Targeted insertionvia
`P3(FE35035)
` ae
`
`homologous recombination9“yn ”
`LAK
`20K
`
`Note: Drawing not to scale
`
`Fig. 10
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet12 of 13
`
`Site-directed nuclease cut(s}
`
`:
`
`|RBESnnnen~nnan ges~MIR604transgeniclocus ToTFatty
`
`$t$pnCPM
`casera
`ARRONKERARALAEOARAtoSEENSRW RA ARS:
`
`
`
`.
`
`
`
`-AWWANANAMANAARIVARARATANAAAAARARABARARAAAAAN,
`
`IC Exp. Cass,
`SAAAAEAAAARASAGEAAAAHASAISANABAENAALMA
`
`Beececeeeedtae
` PAT Exp, Cass.
`Percerercery
`
`Donor DNA
`
`LEGEPPEEEOLOEEEEEEUEEELOEEEEEEEOUEEEE,
`
`Targeted insertion via
`homologous recombination
`
`eeunbtnnaneitinahhnaahinehAARNEnNNNAAN
`
`—
`
`steeeneneneseennaneeaneneenenen
`
`asenaGRUGAGREARAEAA
`
`
`
`
` mibp.Casszpores
`PrMTL | mCn3A |to _t Exp, Cass,
`
`
`ARAN
`
`ay aad
`
`sannannaanneannnennnn! -sonnannnnnnsnnnnnnntSOR
`
`Targeted recombinant event
`
`Note: Drawing not to scale
`
`Fig, 14
`
`
`
`ATTY DOCKET NO.: 80484-US-REG-D-NAT-2
`
`Replacement Sheet
`
`Sheet13 of 13
`
`“klte-Cirected nuclease cutis) MN
`tt,4Naty DS RR ARE RRA RES
`+ _'
`i
`_pasencecnsuensentnncenesensuencea eeeeneTCHGUESSEREERE
`$
`{BFS
`ytiyt
`
`
`MiR604 transgenic locus
`
`Sy
`
`ihoee aRY RAE ARH RAE AE
`
`AWAY ANAKEAE AY >
`‘\
`we
`
`
`
` Donor DNA
`
`Targeted integration via
`+ homologous recombination
`
`
`
`Targeted recombinant event
`
`Note: Drawing not to scale
`
`Fig. 12
`
`
Accessing this document will incur an additional charge of $.
After purchase, you can access this document again without charge.
Accept $ Charge
Still Working On It
This document is taking longer than usual to download. This can happen if we need to contact the court directly to obtain the document and their servers are running slowly.
Give it another minute or two to complete, and then try the refresh button.
A few More Minutes ... Still Working
It can take up to 5 minutes for us to download a document if the court servers are running slowly.
Thank you for your continued patience.
This document could not be displayed.
We could not find this document within its docket. Please go back to the docket page and check the link. If that does not work, go back to the docket and refresh it to pull the newest information.
Your account does not support viewing this document.
You need a Paid Account to view this document. Click here to change your account type.
Your account does not support viewing this document.
Set your membership
status to view this document.
With a Docket Alarm membership, you'll
get a whole lot more, including:
- Up-to-date information for this case.
- Email alerts whenever there is an update.
- Full text search for other cases.
- Get email alerts whenever a new case matches your search.
One Moment Please
The filing “” is large (MB) and is being downloaded.
Please refresh this page in a few minutes to see if the filing has been downloaded. The filing will also be emailed to you when the download completes.
Your document is on its way!
If you do not receive the document in five minutes, contact support at support@docketalarm.com.
Sealed Document
We are unable to display this document, it may be under a court ordered seal.
If you have proper credentials to access the file, you may proceed directly to the court's system using your government issued username and password.
Access Government Site
