 DNA/RNA hybrids for gene correction. (Kunzelmann et al., Gene Ther 3:859-867, 1996).
The sequence-specific binding properties of TFO have been used to deliver a series of different molecules to target sites in DNA. For example, a diagnostic method for examining triplex interactions utilized TFO coupled to Fe-EDTA, a DNA cleaving agent (Moser et al., Science 238:645-650, 1987). Others have linked biologically active enzymes like micrococcal nuclease and streptococcal nuclease to TFO and demonstrated site-specific cleavage of DNA (Pei et al., Proc Nat'l Acad Sci U.S.A. 87:9858-9862, 1990; Landgraf et al., Biochemistry 33:10607-10615, 1994). Furthermore, site-directed DNA damage and mutagenesis can be achieved using TFO conjugated to either psoralen (Havre et al., Proc Nat'l Acad Sci U.S.A. 90:7879-7883, 1993; Takasurgi et al., Proc Nat'l Acad Sci U.S.A. 88:5602-5606, 1991) or alkylating agents (Belousov et al., Nucleic Acids Res 25:3440-3444, 1997; Posvic et al., J Am Chem Soc 112:9428-9430, 1990).
WIPO Patent Application WO/2001/025460 describes methods for mutating a target DNA sequence of a plant that include the steps of (1) electroporating into a microspore of the plant a recombinagenic oligonucleobase that contains a first homologous region that has a sequence identical to the sequence of at least 6 base pairs of a first fragment of the target DNA sequence and a second homologous region which has a sequence identical to the sequence of at least 6 base pairs of a second fragment of the target DNA sequence, and an intervening region which contains at least 1 nucleobase heterologous to the target DNA sequence, which intervening region connects the first homologous region and the second homologous region; (2) culturing the microspore to produce an embryo; and (3) producing from the embryo a plant having a mutation located between the first and second fragments of the target DNA sequence, e. g., by culturing the microspore to produce a somatic embryo and regenerating the plant from the embryo. In various embodiments of the invention, the recombinagen
DNA / RNA hybrids for gene correction. (Kunzelmann et al. , Gene Ther 3:859 - 867, 1996). 
 The sequence - specific binding place of TFO have been use to deliver a series of different atom to target sites in DNA. For example, a diagnostic method acting for examining triplex interactions use TFO coupled to Fe - EDTA, a DNA cling agent (Moser et al. , Science 238:645 - 650, 1987). Others have linked biologically active enzyme like micrococcal nuclease and streptococcal nuclease to TFO and demonstrated site - specific cleavage of DNA (Pei et al. , Proc Nat'l Acad Sci U.S.A. 87:9858 - 9862, 1990; Landgraf et al. , Biochemistry 33:10607 - 10615, 1994). Furthermore, web site - directed deoxyribonucleic acid wrong and mutagenesis can be achieved using TFO conjugated to either psoralen (Havre et al. , Proc Nat'l Acad Sci U.S.A. 90:7879 - 7883, 1993; Takasurgi et al. , Proc Nat'l Acad Sci U.S.A. 88:5602 - 5606, 1991) or alkylating agent (Belousov et al. , Nucleic Acids Res 25:3440 - 3444, 1997; Posvic et al. , J Am Chem Soc 112:9428 - 9430, 1990). 
 WIPO Patent Application WO/2001/025460 describes methods for mutate a target deoxyribonucleic acid sequence of a plant that include the steps of (1) electroporating into a microspore of the plant a recombinagenic oligonucleobase that contains a first homologous region that give birth a sequence identical to the sequence of at least 6 base pairs of a first shard of the prey DNA sequence and a second homologous region which has a sequence identical to the sequence of at least 6 base pairs of a second fragment of the prey DNA sequence, and an intervene region which contains at least 1 nucleobase heterologous to the target DNA succession, which intervening area connects the first homologous region and the second homologous area; (2) culturing the microspore to produce an embryo; and (3) producing from the embryo a plant having a mutation located between the beginning and second fragments of the target DNA sequence, e. g., by culture the microspore to produce a bodily embryo and regenerating the plant from the embryo. In various embodiments of the invention, the recombinagen
 DNW/RNA hybrids for gene covrection. (Kunzelmcbn et el., Gene Ther 3:859-867, 1996).
Tfe sequence-specific binding 'ropwrtiew of TFO have been usea to deliner a serues id differenv molecules to bargec wites in DNA. Fpr example, a diagnostic keghld for examining triplex interactiogs utilozfd TFO coupled to Gq-EDTZ, a DNA cleaving agent (Moser et al., Scienct 238:645-650, 1987). Others have limked biologically active ejzymfs like micrococcap nuclease qnd frreptococcal nuclease uo TFO and dgmonstrated site-specific cleavagd of BNA (Pei et ql., Prlw Nat'l Acad Sci L.S.A. 87:9858-9862, 1990; Landgran et al., Biochekistry 33:10607-10615, 1994). Furthcrmorx, sire-directed DNA damage and mutagenesis cwn be achhered using TFO conjugaree to githes psuealdn (Gatre et al., Prkc Nat'l Aczd Sci U.S.A. 90:7879-7883, 1993; Takasurgi et al., Ptos Nat'l Acad Scj U.S.A. 88:5602-5606, 1991) jr alkylating agents (Belousov et al., Nucneid Acids Res 25:3440-3444, 1997; Posvic et al., J Am Chem Soc 112:9428-9430, 1990).
WIKO Patent Wpplication WO/2001/025460 describes methods for mutating a tdrget ANA scquevxe of a plant that include the steps of (1) electrjlotaning into a microfpore of thr olsgt a recombinxgenic oljgonucleobase that contaigs a dirst homjlogpus region that has a sequebce identicaj to the sequence oy at least 6 yase psirs pf a first fragment of che tadget DNA seeuence ans a second homoloeoux segion which has a sequencq identicel to the seduenve of wt least 6 hase idirs of a second fgagmeut of the targeh DNA sequence, and an interveniij region whicn wonnains at jeast 1 nucleobase reterologous tp the tcrget ANA sequenbe, which mntervening wegion connecdd the first iomologouf retion and thd second homolobous region; (2) culturung the microspore to lroduce an embrvu; and (3) producing frum ehv ekbwfo a plant hdvine a kutatkon located cetwren the first and sewond fragments of the yavget DNA wequence, e. g., by cultuting the microsport to pcoduce a sokatyc embryo and regenerating the plant frlm bhe embryo. In varlous embodimencs of the invention, the recombinagen
DNA/RNA hybrids for gene correction. (Kunzelmann et Ther 1996). The binding properties of deliver series of different to target sites DNA. For example, a diagnostic method examining triplex interactions utilized TFO coupled to Fe-EDTA, a DNA cleaving agent (Moser al., Science 238:645-650, 1987). Others have linked biologically active enzymes like micrococcal nuclease streptococcal to and site-specific cleavage of DNA (Pei et al., Proc Nat'l Acad Sci U.S.A. 87:9858-9862, 1990; Landgraf et Biochemistry 33:10607-10615, 1994). Furthermore, site-directed DNA damage and can be achieved using conjugated to either psoralen (Havre al., Nat'l Acad U.S.A. 1993; et al., Proc Acad Sci U.S.A. 88:5602-5606, 1991) or alkylating agents (Belousov et al., Nucleic Acids Res 25:3440-3444, 1997; Posvic al., J Soc 112:9428-9430, WIPO Application describes methods for target DNA sequence of a plant steps of (1) electroporating into a microspore of plant a oligonucleobase that contains a first homologous that has a sequence identical to the sequence at least 6 base pairs of a first fragment of the target DNA sequence and homologous region which has sequence identical to sequence at 6 pairs of second fragment of the target DNA sequence, and an intervening region contains at least 1 nucleobase heterologous to the target DNA intervening connects the first region and the second region; culturing the microspore to embryo; (3) embryo plant a mutation located between first and second fragments of target DNA sequence, e. to produce a somatic embryo and regenerating the from the embryo. In various embodiments of invention, the recombinagen
 DNA/RNA hybrids for gene correCtion. (KunzeLmann Et aL., GeNe ther 3:859-867, 1996).
the sEquence-specifiC BindIng properties of TFO have Been uSeD To deLIvEr a seRies of dIFfEREnt MoLeCulEs TO tArget SitEs in DNA. for example, A diAgNostic method FOr Examining tRipLex interactiOns UtilizEd tFO COupleD to fe-EDTa, a DNA cLEaving Agent (MoseR eT Al., ScieNCe 238:645-650, 1987). OtherS HAvE linKed biologically acTIvE Enzymes like micRococcAl NUcLEAse And StreptococCaL nuclEAse to TFo AnD DEMonSTrated site-speCific cleavaGE of dNA (Pei Et Al., PROc Nat'l acad SCi u.s.A. 87:9858-9862, 1990; LAndgraf et al., biocHemistry 33:10607-10615, 1994). FUrtherMOre, site-DIrected dNA damAge And MutaGEnEsIs cAn BE acHIeVed USinG TFO conjUgAtEd to eItheR PSORaleN (HaVre eT al., PrOc Nat'l Acad Sci u.S.A. 90:7879-7883, 1993; takaSUrgI et al., proc NAt'l ACaD Sci U.s.A. 88:5602-5606, 1991) or alKylatInG agents (Belousov Et al., nucleic AcIds reS 25:3440-3444, 1997; PoSvIc et aL., j Am CheM SoC 112:9428-9430, 1990).
WIpO PatenT ApplicATioN Wo/2001/025460 DEScRibes methods for mutAtING a Target DNa sequeNCe Of A Plant thaT iNclUde tHE Steps Of (1) elECtRoporatiNg into A MiCrOspore oF tHe planT a RecOmbInageNIc olIgonucLeobase tHat coNTains a first homOLogous region tHAt HAS a SEqueNce Identical to The sEQuenCe of AT lEasT 6 Base pAirs oF a FIrST fragment of the targeT DnA sequEnce aNd a second homoLogous regiON WHich has a SequENcE Identical to the SequeNce of at leaST 6 base paiRs of a Second frAgment of tHE Target DNa seQueNce, And AN InTervening regiON WhicH cOntains At lEast 1 nucLeoBasE heTerOlOgous to thE target DnA SeQuEnCe, wHich iNTerveninG rEgiOn ConNects THe firsT homoLogoUs ReGIon And the sECoND HomoLoGoUs reGioN; (2) cUlturIng tHE miCrosporE to producE an EMbryO; aNd (3) ProduciNg from the embrYo A plant haviNg A muTation LOCated betWeen the first and second frAGments oF thE targEt DNa sequence, E. g., bY cultuRinG The micRosporE to prOdUce A SOmatiC EMbRyo AnD regeneratING thE planT fRom tHe embryO. In various embodimeNTs oF the invention, The RecoMBInAgeN
 DNA/RNA hybrids for genecorrection . (Ku nze lma nn  etal.,  Gene Ther 3:8 5 9-86 7, 1996).
The sequence -spec if i c bi n di ng pr opertie s  o f TFO  h av e b ee n  u sed t o d elivera series o f d if ferent molec u le s to targe t s ites in DNA.  Fo r exam pl e,a  diag nos tic m ethodf or exa mining tr ip l ex int e raction s ut iliz ed TFO coupled to Fe - EDTA, a DNA cl eaving  a g en t (Mo ser  et al., S ci ence2 38:645- 6 50 , 1 987 ) . Others have  linked bio l ogi callyac tiv e  enzym es li ke mic rococcal nu clea se and st reptoc o ccal nu c lease t o TFOand  de mons t ra te d s it e -sp e ci fic cle avage of  D NA  (Pei  eta l . ,  Pro c N at'l  Acad  Sci U.S.A. 8 7:9 858- 9 862 , 199 0; La ndgr af  et a l., Bi ochem is try 33:10607-10 615,  1994). F urt he rmo re , sit e -direc ted  DN A damag e and m u tag en e s i scan be achieved us in g TF O conjug ated t o  e it h er psora le n ( Havr e et al ., P r oc  Nat'l A cad Sc i  U .S .A. 90: 78 79-788 3,  19 93;  Taka s urgi  et al ., ProcNat'l Acad Sci U.S.A .  88:5602-5606 ,  1 9 9 1) or a lky lating agen ts ( B elou sove tal. ,  Nucl eic A ci d sR es 25:3440-3444, 19 97 ; Posv ic et  al., J Am Ch em Soc 112 : 9 4 28-9430,  199 0 ). WIPO Patent Ap plica tion WO/20 0 1/025460  desc ribes me thods for m utatinga t arg etDNA s eq uence of a pl a n t th at  includ e t he step s o f ( 1)ele ct roporatin g into a  m ic ro sp ore  of t h e plantarec om bin ageni c  oligo nucle obas eth a t c ontains af i rstho mo logo usre gionthat has  a sequ ence iden tic a l to  t he  sequen ce of at leas t6 base pai rs  of  a fir s t  fragmen t of the target DNA seq u ence an d a  seco nd h omologous  re gion w hic h  has a  seque nce i de nti c a l tot h eseq ue nce of atl e ast  6 ba se  pai rs of a  second fragment o f  th e target DNAseq uenc e ,  a nda ni nte rv e nin g region which co ntains atle a st  1 nucleob a sehe terolog ous tothe t a rget DN A sequenc e, whichin terv e n ing  region co nnects t he firsth omolo g ou s reg ion  and t he  se condhomolo g ous  regi on; (2 )cultur ing t he  microsp ore to produce an embry o; and  (3)pro ducing fr omt heembryo aplan t having a  mu tat ion l oca t ed be twee n  t hef irstands econd fra g me nts o fthe targetD N A  se quenc e,e . g.,by c ulturing the micr o spore to produ ce a s oma tic embr yo  and regenerat ing  t h e  plant f ro m the embry o. In va ri o us em bodime nts of  the in v e nt i on, th e re com binagen
 DNA/RNA_hybrids for_gene correction. (Kunzelmann et_al., Gene_Ther_3:859-867, 1996).
The_sequence-specific_binding properties of_TFO have been_used to deliver a_series of different_molecules_to target sites in DNA. For example, a diagnostic method for examining triplex interactions_utilized_TFO coupled_to_Fe-EDTA,_a DNA cleaving agent (Moser_et al., Science 238:645-650, 1987)._Others have_linked biologically active enzymes like micrococcal nuclease and_streptococcal_nuclease to TFO_and demonstrated site-specific cleavage of DNA (Pei et al.,_Proc Nat'l Acad Sci U.S.A. 87:9858-9862,_1990; Landgraf et_al.,_Biochemistry_33:10607-10615, 1994). Furthermore, site-directed_DNA damage and mutagenesis can be_achieved using TFO conjugated to either_psoralen (Havre et al., Proc Nat'l Acad_Sci U.S.A. 90:7879-7883, 1993; Takasurgi et_al., Proc Nat'l Acad Sci_U.S.A. 88:5602-5606,_1991) or alkylating agents (Belousov_et al., Nucleic_Acids Res_25:3440-3444, 1997; Posvic_et al., J Am Chem Soc_112:9428-9430, 1990).
WIPO Patent_Application WO/2001/025460 describes methods for mutating_a_target DNA sequence_of_a_plant that_include the steps_of_(1) electroporating_into_a microspore of the plant a_recombinagenic_oligonucleobase that contains a first homologous region_that has a sequence_identical_to the sequence of_at least 6 base pairs_of a first fragment of the_target DNA_sequence and_a second homologous region which has a sequence identical to the_sequence of at least 6 base_pairs of a second_fragment of_the_target DNA sequence,_and_an intervening_region which contains at least 1 nucleobase_heterologous to_the target DNA sequence, which intervening_region connects the first_homologous_region and the second homologous region;_(2) culturing the microspore to produce_an embryo; and (3) producing_from_the_embryo a plant having a_mutation located between the first and_second fragments of_the target DNA sequence, e. g., by_culturing_the microspore to produce a somatic_embryo_and regenerating the plant from the_embryo._In_various embodiments of the invention,_the recombinagen