Large-scale discovery of induced point mutations with high-throughput TILLING.
TILLING (Targeting Induced Local Lesions in Genomes) is a general reverse-genetic strategy that provides an allelic series of induced point mutations in genes of interest. High-throughput TILLING allows the rapid and low-cost discovery of induced point mutations in populations of chemically mutagenized individuals. As chemical mutagenesis is widely applicable and mutation detection for TILLING is dependent only on sufficient yield of PCR products, TILLING can be applied to most organisms. We have developed TILLING as a service to the Arabidopsis community known as the Arabidopsis TILLING Project (ATP). Our goal is to rapidly deliver allelic series of ethylmethanesulfonate-induced mutations in target 1-kb loci requested by the international research community. In the first year of public operation, ATP has discovered, sequenced, and delivered >1000 mutations in >100 genes ordered by Arabidopsis researchers. The tools and methodologies described here can be adapted to create similar facilities for other organisms.
Till
Bradley J
BJ
Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
Reynolds
Steven H
SH
Greene
Elizabeth A
EA
Codomo
Christine A
CA
Enns
Linda C
LC
Johnson
Jessica E
JE
Burtner
Chris
C
Odden
Anthony R
AR
Young
Kim
K
Taylor
Nicholas E
NE
Henikoff
Jorja G
JG
Comai
Luca
L
Henikoff
Steven
S
D000477:Alkylating Agents / Q000009:adverse effects; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000295:instrumentation* / Q000379:methods*; D018744:DNA, Plant / Q000235:genetics / Q000378:metabolism; D005038:Ethylnitrosourea / Q000009:adverse effects / Q000031:analogs & derivatives; D017343:Genes, Plant / Q000187:drug effects / Q000235:genetics; D018745:Genome, Plant; D020407:Internet; D016296:Mutagenesis / Q000187:drug effects / Q000235:genetics; D017354:Point Mutation / Q000187:drug effects / Q000235:genetics*; D012984:Software
D000477:Alkylating Agents; D018744:DNA, Plant; D005038:Ethylnitrosourea