Antisense inhibition of beta-glucuronidase expression in preimplantation mouse embryos: a comparison of transgenes and oligodeoxynucleotides.
Antisense inhibition of beta-glucuronidase expression in preimplantation mouse embryos: a comparison of transgenes and oligodeoxynucleotides.
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植入前小鼠胚胎中β-葡萄糖醛酸酶表达的反义抑制：转基因和寡脱氧核苷酸的比较。
DOI：
10.1089/ard.1991.1.1
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Karolyi,J
中科院分区：
文献类型：
--
作者：
Ao,A;Erickson,RP;Bevilacqua,A;Karolyi,J
我们以植入前胚胎中β-葡萄糖醛酸酶表达的抑制为模型，将引导反义RNA和反义寡脱氧核苷酸合成的转基因注射与我们之前的细胞质注射反义RNA的结果进行了比较。将含有1.4 kb β-葡糖醛酸酶编码区的反义DNA原核注入小鼠金属硫蛋白I启动子，可短暂抑制植入前小鼠胚胎的基因表达。原核注射较小的反义DNA结构，重叠起始密码子，未能抑制基因表达。注射两个20-mer反义寡脱氧核苷酸，一个与起始密码子序列互补，另一个与β-葡萄糖醛酸酶基因外显子7序列互补，未能抑制基因表达。此外，胚胎培养中存在反义寡脱氧核苷酸对基因活性没有影响。使用放射性标记的低聚物添加到培养基中，我们发现胚胎对低聚脱氧核苷酸的摄取很差。
Using as a model the inhibition of β-glucuronidase expression in preimplantation embryos, we have compared injections of transgenes directing the synthesis of antisense RNA and antisense oligodeoxynucleotides to our previous results with cytoplasmic injections of antisense RNAs. Pronuclear injection of an antisense DNA construct containing 1.4 kb of coding region of β-glucuronidase fused to the mouse metallothionein I promoter results in transient inhibition of gene expression in preimplantation mouse embryos. Pronuclear injection of a smaller antisense DNA construct, overlapping the start codon, failed to inhibit gene expression. Injection of two 20-mer antisense oligodeoxynucleotides, one complementary to sequences including the initiation codon and the second complementary to exon 7 sequences of the β-glucuronidase gene, failed to inhibit gene expression. In addition, cultures of embryos in the presence of antisense oligodeoxynucleotides had no effect on gene activity. Using radiolabeled oligomers added to the culture medium, we found poor uptake of oligodeoxynucleotides by embryos.