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反义寡脱氧核糖核苷酸介导的非洲爪蟾卵母细胞和胚胎中母源mRNA的切割

Antisense oligodeoxyribonucleotide-directed cleavage of maternal mRNA in Xenopus oocytes and embryos.

作者信息

Shuttleworth J, Matthews G, Dale L, Baker C, Colman A

机构信息

Biochemistry Department, University of Birmingham, U.K.

出版信息

Gene. 1988 Dec 10;72(1-2):267-75. doi: 10.1016/0378-1119(88)90152-7.

Abstract

We have investigated the effect of specific antisense oligodeoxynucleotides (oligos) on endogenous histone H4 mRNA in Xenopus oocytes, eggs and embryos. In unfertilised eggs and non-matured oocytes, one 20-mer oligo (H4-1) mediated the RNAse H-like cleavage of up to 95% of H4 mRNA (which included polysomal mRNA), and cleavage was still obtained when the size of the oligo was reduced to a 10-mer; no cleavage was observed with 6- and 8-mers. The residual uncleaved mRNA appeared to be completely inaccessible to H4-1 since a second injection caused no further cleavage. A second 20-mer (H4-2) directed against a different region of H4 mRNA was much less effective (less than 5% cleavage). In fertilised embryos, injections of H4-1 and an oligo directed against the localised Vg1 mRNA caused less cleavage than in oocytes and also showed signs of inducing localised, non-specific mRNA cleavage. However we have been able to prepare fertilised embryos devoid of Vg1 mRNA by maturing and fertilising oligo-injected oocytes in vitro.

摘要

我们研究了特异性反义寡脱氧核苷酸(oligos)对非洲爪蟾卵母细胞、卵和胚胎中内源性组蛋白H4 mRNA的影响。在未受精卵和未成熟卵母细胞中,一种20聚体寡核苷酸(H4-1)介导了高达95%的H4 mRNA(包括多聚核糖体mRNA)的类似RNA酶H的切割,当寡核苷酸大小降至10聚体时仍能获得切割;6聚体和8聚体未观察到切割。剩余未切割的mRNA似乎对H4-1完全不可及,因为第二次注射未引起进一步切割。针对H4 mRNA不同区域的第二种20聚体(H4-2)效果要差得多(切割率低于5%)。在受精胚胎中,注射H4-1和针对定位的Vg1 mRNA的寡核苷酸引起的切割比在卵母细胞中少,并且还显示出诱导定位的、非特异性mRNA切割的迹象。然而,我们已经能够通过在体外使注射寡核苷酸的卵母细胞成熟和受精来制备不含Vg1 mRNA的受精胚胎。

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