KHOA HOC CONG NGHE
HIEIV T R A I V G COIXIG M G H E C H I I X I H S I J A GEIM C R I S P R / C a s g T R O I V G C A I TIEIXI M O T S O TilXIH TRAIXIG O
C A Y L L I A V A TRIEIXI VOIXIG O V I E T IXIAIVI
Nguyen Ti6n DOng', La Van Hi^n^
T6MTAT
Phat trien cay trong dot bien thong qua chinh sua gen la hircmg di day tnen vong va linh hoat trong cong tac chpn giong cay trong phuc vu cho mot nen san xuat nong nghiep ben vimg. Trong do, nghien ciiu chinh siia he gen bang CR1SPR/Cas9 tao ra nhimg thay doi 6 miic do phan tu va dam bao tinh dac hieu. Ky thuat nay nhanh chong, don gian va hieu qua cao hon so voi cac phuong phap gay dot bien thong thuong nhu ZFNs va TALENs. Thiet ke vecta CRISPR/Cas9 danh rieng cho nhieu loai thuc vat da duoc thiet lap, ap dung thanh cong tren cay mo hinh Arabidopsis tbaliana va cac cay luong thuc quan trong nhu lua, dau tuong, ngo. Chinh siia he gen bang CRISPR/Cas9 se tao ra nguon vat lieu co gia tri cho cong tac chon gidng dpt bi^n. Bai viet nay torn tat vai tro cua cong nghe CRISPR/C3s9 trong chinh sua he gen va moi sd thanh tuu dat duofc cung nha tiem nang ung dung trong cai thien di truydn cay liia lien quan d^n nang suiit, ch^t luong va kha nang chdng chiu cac dieu kien bat lm ciia moi trudng.
Tir khoi CRISPR/Cas9, dot bie'n, GMO, Itia, stress.
I.HETHdniGCRISPR/CasS
CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeat/Cas9) duac phat hi$n trong he thdng miln dich ciia vi khuan chong lai su xam nhap cua DNA (Deoxyribonucleic acid) ngoai lai, nhu thuc khuan the hay plasmid. Khac voi he thong miln dich dua tren cac enzyme cat gioi han, he thong nay pha huy DNA ngoai lai bang phan tu RNA.
He thong bao gom trinh tu lap CRISPR va mot protein Cas9. Viing CRISPR bao gom su ket hpp cua gen Cas9, trinh tu khong ma hoa (non-coding) RNA gpi la CRISPR RNA (crRNA) va trinh tu ngdn chuyen ma (trans-activating; tracrRNA) (Barrangou, 2013;
Bortesi va Fischer. 2015). Hai trinh tir crRNA va tracrRNA dupc bi^t den nhu mot RNA d&n duong (guide RNA, gRNA). no xac dinh tinh dac hieu vi tri phin chia nucleotit muc tieu thugc motif li^n ke tien vimg d6m (Protospacer Adjacent Motif -PAM), trinh tu PAM gom 3 nucleotitde 5'-NGG-3' nam o phia sau (downstream) cua DNA muc tieu, la dieu kien dau tien cho DNA tach doi boi Cas9. Trong do, 20 nucleotit xac dmh vi tri dac hieu, goi la hat nhan xap xi 12 nucleotit (vang cam) nam phia truoc (upstream) cua PAM la dac biet quan trpng dpi voi
qua trinh bat cap giira RNA va DNA muc tieu (Hinh 1) (Barrangou, 2013; Jinek va cs., 2012). Protein Cas9 la mot enzyme endonuclease g6m hai tieu phdn RuvC va HNH, m6i tieu phan cat mpt chu6i DNA (Hinh 1). Enzyme nay h6 tro CRISPR tai vi tri spi doi DNA bi pha vo (double-straded breaks, DSBs) boi mot trinh tu gRNA g6m khoang 20 nucleotit bo sung cho doan DNA muc tieu (Cong va cs., 2013). Mot protein Cas9 co kha nang su dung cho nhilu gRNA khac nhau huong den nhilu vi tri muc tieu trong bo gen Qinek va cs., 2012; Jiang va cs., 2013; Wang va cs.,2016).
So voi ky thuat chinh sua he gen dua tren hoat dong ciia enzyme endonuclease nhu zinc finger nucleases (ZFNs) va transcnption activator-like effector nuclease (TALENs), ky thuat chinh sua he gen CRISPR/Cas9 dupc ung dung rong rai hon do co nhieu ITU diem vupt trpi nhu don gian, hieu qua dot bien cao. Vi du, tren cay ngo, CRISPR/Cas9 va TALENs gay dot bien o cung vi tri muc tieu dat hieu qua lan lugt la 13.1% va 9,1% (Liang va cs., 2014). Tuy nhien, de dat dupc kit qua nay con phu thuoc vao trinh tir muc tieu, cau tnic va trinh tu gRNA, protein Cas9 (codon toi uu), chiln luoc bilu hien gRNA va dp nhay cua phuong phap phan tich (Shan va cs., 2013; Bortesi va Fischer, 2015).
Khoa Cong nghe Sinh hoc va Cong nghe Thuc pham, Trucrng Dai hoc Nong Lam, Dai hoc Thai Nguyen Email: [email protected]
KHOA HOC C b N G NGHE
Hinh 1. Cau true ciia CRISPR/Cas9 va RNA-dan DNA t4ch d6i boi Cas9
(a) O hi thdng tu nhien, Cas9 protein (xanh nhat) duoe din bing mot cau tine hmh thinh bdi CRISPR RNA (crRNA, mau den), bao gdm 20 nucleotit xac dinh vi tri die hiiu vi mdt ydu td chuyen ma CRISPR RNA (tracrRNA, dd), dim bao dn dinh ve eau true vi hoat dong cua Cas9 de phan tach DNA muc tiiu protospacer) vi suhlin dien cua motprotospacer-adjacentmotif (PAM, ving). (b) Cas9cd the duoc lap trinb Ial detach DNA boi mot phin tif RNA din sai don (gRNA, xanh la ciy), mdt DNA hoin ehinh (chimera) duoc tao ra bing cich kit hop dau 3' eua crRNA vdi diu 5' cua tracrRNA (Boriesi vi Fischer, 2015).
2 . UNG DUNG CRISPR/CasS TRONG CAI TBU MOT SO TIUH TRANG 0 CAY LUA
2.1. Tinh hinh nghidn cijfu trfin th^ gi6i Cho den nay, he thong chinh
khac nhau Qiang va cs., 2013; Shan va cs., 2013), trong do cay liia duoc nhilu nha khoa hgc quan tam cai thien cac dac tinh nong sinh hpc quan trpng bao gom cai thien nang suat, chat luong, tinh khang cac stress sinh hpc va phi sinh hoc (Bang 1).
sua gen CRISPR/Cas9 da dugc ap dung tren 20 loai cay trong
Bang 1. CRISHt/Cas 9 chinh s6a gen lifen quan d^n cAc tinh trang dugc c&i t h i ^ a cay liia Dac diem Gene muc tieu Ky thuat ap
dung Chtic nang Tham khao
Cai tien nang suat va chat lugng
GW2, GW5, TGW6
CRISPR/Cas9 Tang khoi lugng hat Xu va cs., 2016; Su va cs. 2011
Gnla, GS3
DEPl, CRISPR/Cas9 Tang s6 lugng hat, kich thuoc hat
Li M va cs., 2016; Xu va cs.
2016 . TGW2, GSE5,
GSNl, GSW6
CRISPR/Cas9 Tang chilu dai hat Su va cs., 2011; Guo va cs.
2018 PYLs CRISPR/Cas9 Cai thien tang trudng
va nang suat
Miao va cs., 2018
BADH2 CRISPR/Cas9 Tang huong thom Shan vacs., 2015 Khang stress
sinh hoc
OsSWEETlS/
14
TALENs Tang cuong kha nang khang benh bac la
Antony va cs., 2010; Li va cs., 2012; Blanvillain-Baufum va cs. 2017
TALENs Tang cuong kha nang khang benh bac la
Cai va cs., 2017
OsER922 CRISPR/Cas9 Tang cuong kha nang khang benh dao on
Wang va cs., 2016
Khang stress phi sinh hoc
BEU ALS, OsEPSPS
CRISPR/Cas9 Khang thuoc diet c6/glyphosate
Sun va cs., 2016
ARG0S8 CRISPR/Cas9 Khang han Shi vacs., 2017 Cai thien dinh
duong
SBEllb, SBEI CRISPR/Cas9 Tang ham lugng linh hot
Sun va cs., 2017
NONG NGHIEP VA PHAT TRIEN NONG THON - KY 1 - THANG 7/2020
KHOA HOC CONG NGHE
2.1.1. Cai thiin nang sui't va chit luang hat O cay Ilia nang suat dugc quy dmh boi hai hoac nhilu gen (Xing va Zhang, 2010). Nhieu nhom nghidn curu da co gdng thilt lap ban do cac tinh trang so lugng (locus trait quantative- QTL). Cac gene tilm nang co the duoc chuyen true tilp hoac gian tiep vao cay de phat trien cac giong co nang suat cao (Shen va cs., 2018). Tuy nhien, phuong phap nay rat kho ddi voi cac tinh trang dugc quy dinh boi nhieu gen va kho ki^m soat duoc vi tri cua cac gen muc tieu. Khac voi phuong phap tren, CRISPR/Cas9 co kha nang nhan dien chinh xac DNA muc tieu bang phan tu gRNA va chinh sua bdng enzyme Cas9 (Cong va cs., 2013; Lin va cs., 2014; Bortesi va Fischer, 2015).
Bang ky thuat nay, tinh trang hat o cay lua da dugc cai thifin thong qua chinh sua trinh tu gen kilm soat kich thuoc hat GS3 (Grain size 3) va so lugng hat tren bong Gnla (Gram number la) (Li va cs., 2016;
Shen vacs., 2018).
Nghien cuu cua Xu va dong tac gia (2016) cho thay nang suat liia da dugc cai thifin sau khi loai bo cac gen lien quan den kich thuoc, so luong va Idioi lugng hat nhu grain size 3, 5 (GS3, GS5), dense and erectpanicle 1 (DEPf), gram weight2 {GW2), grain number la (Gnla). Li va cpng su (2016) da chinh sua bon gen rieng rfi Gnla, DEPl, GSSva IPAI thu dugc cac cay liia co kieu hinh nhu so hat/bong tang, nhilu hat chdc/bong va kich thuoc hat Ion hon so voi cay doi chung. 6 mot nghifin cuu khac, Xu va cpng su (2016) cho rang dot bien d i n g thoi ba gen hen quan din khoi luong hat GW2, GW5 va thousand grain weight 6 (TGWd) co the lam tang 29,3% khoi luong hat so voi ban ddu. Dilu nay cho thay CRISPR/Cas9 CO kha nang gay dot biln cac gen lifin quan din nang suat Ma.
Ngoai ra, CRISPR/Cas9 cung duoc tmg dung d l cai thifin chat luong gao thong qua chinh sua cac gen chi phoi. Vi du nhu gen betaine aldehyde dehydrogenase 2 (BADH2) kiem soat kha nang tong hgp ham lugng 2-acetyl-l-pyrroline (2AP) quy dinh huong thom chinh trong gao. Bang ky thuat CRISPR/Cas9, Shao va cpng su (2017) da chinh sua gen BADH2 lam tang ham hugng 2AP a giong lua indica Zhonghua 11 ddn din huong thom ciia gao tang len dang ke. Sun va cong su (2017) da thanh cong tao ra hat gao co ham luong tinh bot cao bang each chinh sua gen starch branching enzyme (SBE), SBEI va SBEIIb. Trong khi cac dong dot bien gen
Sfif/khong lam tang ham luong tinh bpt, cac dot biln gen SBEIIcho thay ham lugng tinh bpt cao hon 25 ldn so vol doi chimg. Dilu nay cho thay gen SBEIl dong mot vai tro quan trpng trong vific xac dinh cau tnic va tinh chdt a i a tinh bpt cua hat gao.
2.1.2. Cai thien kha ning chdng chiu Stress sinh hpc do cac vi sinh vat gay bfinh bao g6m vi khuan, ndm vi nit va con tning tao ra lam ton thdt toi 42% nang suat Iuong thuc toan cdu (Oerke, 2005). Do vay, phat trien cay trong khang bfinh la nhu cau can thilt, trong do mot so gen lien quan den benh da dugc gay dot bien bang cac phuong phap chinh sua gen de tao ra giong liia khang bfinh.
Zhou va cong su (2015) sii dung ky thuat CRISPR/Cas9 de chinh sua gen OsSWEETlS tren giong Ilia IR24 tao ra cac dong dot bien co kha nang de khang doi voi benh bac la. Wang va cgng su (2016) gay dot bifin gen OsERF922hang each chen hoac xoa vi tri dich tao ra tdn so dot bien len toi 42%
o cac cay TO. Danh gia kieu hinh cua 6 dong dot biln dong hgp tuT2 cho thdy ty le nhilm benh dao on da giam dang ke so voi cac cay doi chung. Gen eukaryotic translation initiation factor 4G {elF4G) duoc cho rdng co ben quan den cac phan ung khang lai vi nit RTSV (Rice tungro spherical virus) gay benh vang lun b liia (Lee va cs., 2010). Dieu nay da dupc chung minh khi Macovei va cpng su (2018) gay dot biln gen elF4G a giong liia IR64 mdn cam voi vi nit RTSV tao ra cdc dong liia co tinh khang cao.
Ngoai cac nghien cuu tang kha nang chong chiu cac stress sinh hpc nhu trfin, cac nha khoa hoc cung chu trpng den vific nang cao kha nang chong chiu cac dieu kien bdt lgi cua ngoai canh (phi sinh hgc).
Gan day, Li va dong tac gia (2016) da thay t h i gen dua tren ky thuat TALEN va tao ra dot bien diem kep a cay liia mang gen ma hoa enzym acetolactate synthase (ALS). Hieu suat dot bien la 6,3% voi toan bp dot bien on dinh cho kha nang khang thudc diet CO manh. Tuong tu, thong qua CRISPR/Cas9 Sun va cpng su (2016) tao ra nhieu dot biln dilm rieng le h-fin trinh tu gen ALS. Ket qua sang lpc kieu hinh cho thdy cac dong chinh sua bieu hifin kha nang khang thudc diet co sau 36 ngay phun. Nang cao kha nang chdng chiu nhiet dp thdp o liia cung dugc nhilu nha khoa hpc quan tam. Huang va cpng su (2017) da chinh sua gen TIFYla va TlFYlb hen quan den kha nang chong chiu nhiet dp 6 cay liia. Cac cay dot bien
KHOA HOC CdNG NGHfe
bieu hien ro ret kha nang chdng chiu vm nhiet do thap cua moi truong.
2.2. Tinh hinh nghiin cuu a Vi^t Nam O nuac ta, cong nghe chinh siia gen CRISPR/Cas9 vdn con kha mod me, dang dugc tiep cdn, va trien khai vao thuc tiln. Hien nay, Cas9 dang dugc mpt sd vien va trung tam tilp thu, hao doi voi ddi tac qudc te trong khuon khd cac de tai, du an hgp tac qudc t l ding nhu cac du an trong nuoc. Chinh sua gen CRISPR/Cas9 buoc dau xay dung va ung dung thanh cong trfin mot sd cay trdng quan trong nhu Ilia, thudc la, va ca chua. Tai Vien Cdng nghe Sinh hpc, nhom nghien ciiu tai Phong Cong nghe t l bao thuc vat da su dung CRISPR/Cas9 thanh cong tao dot bifin gen tren cay dau tuong va thudc la, voi hieu suat tao dot bien dau tuong >90% va 20-40% a thudc la. Ket qua phan tich dot biln cho thdy khong anh huong den kha nang sinh truong, phat trien cila cay dau tirong va thudc la so voi gidng gdc.
Hon nira, chinh sua gen CRISPR-Cas9 la mot trong nhiing huong di moi cua Vien Di truyin va Hgc vien Nong nghifip Viet Nam trong giai doan 2015- 2025. O giai doan nay, chuong trinh cong nghfi sinh hpc nong nghifip "Ung dung cdng nghi chinh sua hi gen de cai tao tinh trang mui thom vi khing binh bac la & mot sd gidng Ida chu Iuc cua Viit Nam "thuc hien tai Vien Di truyin Nong nghifip trong giai doan 2017-2020. Voi ket qua dir kien chinh sua viing promoter cua gen OsSWEET14 tren gidng Bdc Thom 7 va gen BADH2tven gidng liia OM5451. Ben canh do, cac nha khoa hoc cua Hpc vifin Nong nghiep Viet Nam dang tien hanh nghien ciiu su dung CRISPR/Cas9 tao dot biln dinh huong hren gen LiA9 a cay ca chua nhdm tao dong ca chua khong hat.
Nhirng dinh huong va du kien kit qua dat dugc a mpt sd CO quan nghien ciiu cho thay kha nang tiep nhan va lam chii cong nghe chinh sua gen cung nhu tiem nang ting dung cong nghe nay trong chgn tao gidng cay trdng dot bien oViet Nam.
3. TIEM NANG UNG DUNG 0 VIET NAM
Chpn tao gidng liia a nuoc ta chu yeu thong qua lai tao truyin thdng, xu ly dot biln, chgn lpc va tuyIn chon cac dong nhap ngi. Phuong phap nay doi hdi mat rat nhieu thoi gian, chi phi de tao ra gidng moi.
Vi thi, mgt sd nha khoa hoc da iing dung cong nghe moi trong chpn tao gidng lua nhu chpn gidng bdng chi thi phan hi ADN. chuyen gen thong qua cac hop
tac nghien ciiu voi cac nha khoa hgc nuoc ngoai.
Nhung nam gdn day, Vifin Liia ddng bang song Ciiu Long da tao ra nhilu gidng liia moi nho img dung chi thi phan tu MAS frong chgn gidng liia chiu man, chiu nong, khang rdy,... Vifin Di truyen Nong nghiep cung tao ra cac gidng liia thom, chiu iing thong qua chi thi phan hi. Phirong phap nay mac dii mang lai hieu qua nhung vdn mat nhieu thoi gian lai tao va chgn loc.
Mat khac, tao gidng cay trdng moi thong qua chuyen gen cho thay tilm nang irng dung nhanh chong va hieu qua. Ky thuat nay dang dupc iing dung tren dau tuong, ngp va bong. Lgi ich cay hdng bien ddi gen mang lai da dupc da sd nguoi trdng tren t h i gioi thira nhan. Tuy nhien o nuoc ta viec nghien ciiu chuyen gen tren cay liia khong dugc khuyen khich boi lien quan din vdn de xuat khau gao va dSc biet khi van con nhirng lo ngai vl su an toan den siic khoe cua ngudi tifiu dimg. Cong nghe chinh sua h^
gen CRISPR/Cas9 cho phep tao ra cSy hdng moi ma khong cdn chuyen bdt cii mot gen ngoai lai nao vao cay chu vi t h i loai bd hoan toan lo ngai ve gen khang khang sinh Cling nhu cac gen chi thi khac thuong thay a phirong phap chuyin gen. Cong nghe nay co the irng dung trong cong tac chgn tao gidng lua dap irng yeu cdu san xudt o Viet Nam trong dilu kien thuan lgi, kho khan va dinh huong nhu sau:
S.L Thuin lgi
3.1.1. Vai tid va tam quan trong cua viic nghiin cuu nhanh chdng tao ra gidng Ida nang suat cbii Immg/ehdng chiu
Liia gao la mat hang nong san xuat khau chii Iuc cua qudc gia chm anh huong nhilu boi khi hau thay ddi va chat lupng chua cao, dieu nay ddn din gia tri gao tren thi truong xuat khau va thu nhap ciia nong dan trong lua con thap. Thuc t l tren doi hoi phai tao ra gidng liia chdng chiu dirge nhilu ylu td bat lgi cung liic, nhung van dam bao chat luong. Voi phuong phap tao gidng hifin nay thirong chi tao ra gidng khang voi mot vai yeu td bat lgi. Do do, CRISPR/Cas9 voi each tiep can co kha nang tao dot biln da gen cung mpt liic se la co hoi tao ra cac gidng Ilia khang da yeu td can thilt de iing pho voi biln ddi khi hau va dam bao san xuat ben vimg. Hon nira, vai su dac hieu vdn co CRISPR/Cas9 true ulp gay ra nhiing dot bien a cac gen muc neu se la co so d l lua chgn cac dot bien tdt co kha nang tang nang sudt, chat lugng va chdng chm stress.
KHOA HOC CONG N G H £
3.1.2. Vi phdng thi nghlim
De dap ling nhu cdu trong cong tac nghien ciiu va tao dong liia dot bien, he thdng ca so phong thi nghiem dugc thilt lap d- cac vien, trung tam nghien ciiu va truong dai hpc. Bao gdm 1 phong thi nghiem trong diem cong nghe gen (Vifin Cong nghe Sinh hgc) va 2 phong thi nghiem trgng diem cong nghfi t l bao thuc vat thuoc Vien Di truyin Nong nghiep va Vien Sinh hgc Nhiet ddi va nhilu phong thi nghiem sinh hpc phan tu quy mo nhd o cac truong dai hpc khdi nghanh nong - lam nghiep. Thong qua cac chuong trinh va du an hpp tac qudc t l ddu tu trang thiet bi tai Vien Cong nghe Sinh hpc, Vien Di truyen Nong nghiep va Hpc vifin Nong nghifip Vifit Nam phai k l din du an tai h g cila FIRST, JICA...Cu the, voi nguon tai tro cua FIRST, 2 phong thi nghifim (Tm sinh hgc va Sinh hgc phan tu giam dinh gen thuc vat) da dupc ddu tu nang cap. Tai day, cac trang thiet bi sinh hpc phan tii - ky thuat di truyin kha dong bp va ddy dii dap iing dupc yfiu cdu ciia nhilu cong vific nhu sang lpc gen, thiet ke vecto va tao chung Agrobacterium phuc vu chuyen gen.
3.1.3. Vd hi thdng til sinh
Hien nay, quy trinh tai sinh cay liia da dugc nhilu nghien ciiu trong nuoc cong bd, voi vific lira chgn va sang lgc cac gidng co kha nang tai smh cao phuc vu cho chuyen gen. Kha nang tai sinh thanh cong cac gidng liia ban dia va cac gidng nhap ngi da duoc ghi nhan. Theo k i t qua nghien ciiu cua Phan Thi Thu Hien (2012) cho thdy 25/31 gidng liia nuong Viet Nam co kha nang tai sinh, trong do 4 gidng co ty le tai sinh cao gdm IR78878 (70.3%), Khau cong ton (70,3%), Khau toi (70,3%), Khau h a m hom (75,3%).
Mpt nghien ciiu khac cho thay mot sd gidng nhap npi IR64, J02 va Bdc Thom cd kha nang tai sinh va tilp nhan gen kha tdt, dap iing duoc yeu cau cho nghifin ciiu chuyen gen khang han va khang benh hai (Pham Thu Hdng va cs., 2016). Den nay, quy trinh tai sinh mot sd gidng liia japonica (BM9630, NVl, NV2, NV3, J02) va indica (Huong cdm va BC15) da dugc hoan thien (Phan Thi Huong va cs., 2014). Voi nhiing k i t qua nghien ciiu dat dupc sfi tao tiln de cho cong nghe chuyen gen tao dong liia dot bien sau nay.
3.1.4. Vi nhin luc/dieu kiin nghiin cuu vS chuyen gen
Nhilu chuong trinh doi moi cong nghfi qudc gia tap trung hd tro nghien ciiu tao cay liia chuyen gen da duoc xay dung va hoan thien. Cac d l tai cap Nha nuoc va hop tac qudc t l voi Bi (1997-2001), EU (1998- nay) d l 'Tao cay liia chuyin gen chiu ban va chiu man" da duoc triln khai voi chpn lgc gen, thiet k l vector mang gen, va quy trinh chuyin gen tren cay lua thong qua Agrobacterium da duoc xay dung va hoan thien tai Vifin Cong nghe Sinh hgc. Tuong tu, tai Vien Di huyfin Nong nghifip, cac kit qua nghifin ciru phan lap gen va thilt ke vecto bieu hien gen da dugc hen hanh nhdm tao nguon vat lieu di truyin cho cac nghifin ciiu chuyin gen, tao cay hdng biln ddi gen co kha nang chdng chiu voi dilu kien bdt lgi.
Vien da phan lap duoc 19 gen dilu khien tang cuong tmh chiu han phuc vu nghien ciiu tao giong cay trdng chuyen gen chiu han va da xay dung dugc quy trinh chuyen gen vao gidng liia Indica (PBI), Japonica Q02). ngo, dau tuong va thudc la. Hon niia, Vien Di truyfin Nong nghifip la co so true tilp hop tac nghien ciiu voi Trung tam John Innes va Trung tam phan tich he gen Anh, da tiln hanh giai ma thanh cong 36 gidng liia ban dia uu hi ciia Viet Nam trong nam 2013. Tiep do, du an 600 dong/gidng liia ban dia da d l xuat voi Bp Khoa hoc va Cong nghe hd trg trien khai tieu du an "Nang cao nang Iuc nghifin ciiu, lam chii cong nghe genom hgc (Genomics - Assisted Breeding, GAB) va cong nghe chpn gidng ung dung chi thi phan tir (Marker Assisted Backcrossing - MABC) de chpn tao cac gidng liia khang da yeu td ling pho VOI bien ddi khi hau". Do vay, viec triln khai cac du an tren la dip d l ddo tao, nang cao ngudn nhan Iuc hong linh vuc he gen hgc va hn sinh hgc.
Trong do cac nha khoa hoc Viet Nam da tiep can vdi cac phuong phap giai ma hfi gen tien tien, tien tdi chu dong giai trinh tu, tao lap co so dii lieu he gen ddy du cua cac gidng liia Viet Nam. Ddng thai la co hoi de ap dung cong nghe moi CRISPR/Cas9 chgn tao gidng Ilia dot biln gen moi mpt each nhanh chong, hieu qua.
3.2. Kho khan/thach thiic
De dam bao dam an ninh luong thuc va xudt khau nong san bin viing truoc tinh hang bifin ddi khi hau. cong tac nghien ciiu chgn, tao gidng dang dugc dat len hang dau. Viec nhanh chong tao ra nhiing gidng cay hdng dot bifin cd siic chdng chiu dilu kien bat loi smh hpc va phi sinh hpc iing pho vai biln ddi khi hau ciing nhu pha va rao can de tang nang suat.
KHOA HOC C O N G NGHE
san luong luong thuc. doi hdi phai co nhimg thong tin chinh xac v l kieu gen va kieu hinh ciia cac ngudn vat lieu, dac biet la cac ngudn gen ban dia qui. Mat khac, hong quy trinh chpn tao gidng bdng chi thi phan tu, d l xac dinh sir cd mat cua gen quy dinh chdt luong, gen khang sau benh diu su dung nguon gen khang cua nuoc ngoai CRRI, Trung Qudc). Tuy nhien, kieu gen cua gidng ban dia Viet Nam va Trung Qudc co sukhac bifit, ddn den kho khan trong viec tim kiem va sang lgc gen. Trong khi do, cac ngudn gen liia ban dia rat da dang, nhilu ngudn gen chat luong co kha nang khang sau benh tdt nhung chua dugc khai thac hit. Chua co nhirng cong bd khoa hoc v l xac dinh ban dd di truyin cua cac gen chdt lupng, gen khang sau benh, xac dinh thanh phan alen/locus, so sanh su sai khac giira dc alien chat lugng, alien khang cua cac gidng liia ban dia Viet Nam voi cac gidng lua trfin the giai d l phuc vu c6ng tac lai tao gidng.
3.3. Dinh hudng trong thdi gian toi
Tap trung iing dung CRISPR/Cas9 tao dot biln hfin quan den bieu hifin promoter/gen nang sudt Uen quan din tang khdi lugng hat, sd lupng hat, chieu dai hat, sd nhanh/cay va gen khang benh bac la va dao on. Tilp tuc tao dot biln cai thifin ham lupng amylose va lycopene trong gao, la ca so iing dung chinh siia gen tao ra cac dot bien moi gop phan nang cao chat lupng. Bfin canh do, tiem nang ciia CRISPR/Cas9 hong viec tao dot biln da gen lifin quan den cac hoocmon dieu hoa smh trudng va khang stress, nhung vdn dam bao chat lupng. Chpn t^o dupc cac dong liia d6t biln bdng cdng nghe CRISPR/Cas9 dp dung d quy md phong thi nghifim va nha luoi.
4. KETLUAN
CRISPR/Cas9 la cong nghfi chinh sua gen thao tac mgt each dfi dang, dac hieu va da dupc dp dung thanh cong hfin nhifiu gidng cay hdng quan trong.
Cdng nghfi nay se thiic ddy ca hai hudng nghien ciiu CO ban va iing dung, nang cao hieu bilt chiic nang gen va tao tien d l cai thien cac tinh hang mong mudn 6 cay trdng.
Cong nghe CR1SPR/Cas9 co t h i dupc iing dung trong cong tac chpn tap gidng cay hdng d' Viet Nam.
Dac biet hong cdng tac chpn tao gidng liia de nang cao nang suat va chat lugng va kha nang chdng chiu vai cac dieu kien bat loi ciia ngoai canh.
LOI CAM ON
Nhdm tic gia xin chin thanh cam an Bd Khoa hoc vi Cdng nghe da cap kinh phi cho di tii hap tic nghien euu song phuong va da phuong gitia Trudng Bai hoc Qudc gia Kyungpook Han Qudc vi Trudng Dai hoc Nong Lim, Dai hoc Thai Nguyen ma sd HNQT/SPDP/13.19
TAI \m THAM KHAO
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THE CURRENT STATUS OF GENOME EDITING TECHNOLOGY (CRISPR/Cas9) FOR IMPROVING TRAITS IN RICE AND PERSPECTIVE IN VIET NAM
Nguyen Tien Dung, La Van Hien Summary
Development of mutation crops by genentic edition is conventional and an extremely flexibile tool for sustamable agncultural production. Whereas, genome editing (GE) has been modified at the molecular as the nucleotide and ensured by specificity of CRISPR/Cas9. This technique is more quick, simple, and higli efticiency than ZFNs and TALENs. Various plant-specific CRISPR/Cas9 vector systems have been estabilished for adaptation of this technology to many plant species and successfully applied Eo model plant as Arabidopsis thaliana and other important crops such as soybean and com. With this potential powerful and innovative technique, the GE plant breeders make valuale materials for mutation breedmg. In this review, we summanze the role of CRISPR/Cas9 in plant genome editing and current achivements as well as the potential application for nee genetic improvement related to productivity, quality, and tolerant ability to adverse environmental conditions.
Keywords: CmSPR/Cas9, mutants, GMO, nee, stress.
Ngiriri phan bien: PGS.TS. KhuSt Htiu Trung Ngay nhta ba: 13/3/2020
Ng4y thSng qua phan bijn: 15/4/2020 Ngay duyM dang: 22/4/2020
