水稻OsFRK1与OsFRK2基因CRISPR敲除突变体的构建

Generation of CRISPR knock-out mutants of the OsFRK1 and OsFRK2 in rice

为揭示水稻果糖激酶(OsFRK)家族基因的生物学功能,采用CRISPR/Cas9技术,成功创建了2个已鉴定了的水稻OsFRK家族基因的敲除突变体;获得28株OsFRK1的T0代转基因植株,其转基因阳性率、突变率与纯合突变率分别为100%、46.43%和10.71%;获得14株OsFRK2的T0代转基因植株,其转基因阳性率、突变率与纯合突变率分别为92.86%、92.86%和21.43%;T0代敲除纯合突变体中目的蛋白质的功能结构域均发生不同程度的破坏。

To reveal the biological function of fructokinase(OsFRK)family genes in rice,two knock-out mutants for identified OsFRK family genes were constructed using CRISPR/Cas9 technology.The results showed that 28 OsFRK1 and 14 OsFRK2 T0 transgenic plants were successfully constructed.The transgenic positive rate,mutation rate and homozygous mutation rate of OsFRK1 is 100%,46.43%and 10.71%,while OsFRK2 is 92.86%,92.86%and 21.43%.In addition,the functional domains of targeted proteins of the T0 knock-out homozygous mutants were damaged in the mutatnts.