Ribonucleic acid (RNA) interference fragment of zinc finger-x (zfx) gene and application of RNA interference fragment in mouse sex control

Abstract

The invention discloses a ribonucleic acid (RNA) interference fragment of a zinc finger-x (zfx) gene, a carrier formed by the RNA interference fragment and application of the RNA interference fragment in mouse filial generation sex control. The RNA interference fragment has the advantages of hindering, harming or destroying normal development and functions of X-sperms by silencing the zfx gene in an X-sperm of a generative cell of a male mouse. Multiple fertilization chances are provided for a Y-sperm when the male mouse is mated with a female mouse, the sex of a mouse is effectively controlled before fertilization, accordingly the male mouse rate of F1 generation mice is greatly improved, required spermatid is not hurt, and spermatid vitality is not reduced, and the cost is low.

Description

technical field

[0001] The present invention relates to the field of molecular genetics, in particular to the ability to silence zfx RNA interference fragments of genes and their applications. Background technique

[0002] Under natural conditions, the sex ratio of almost all mammals is close to 1:1, thus achieving uninterrupted and stable reproduction of offspring and achieving ecological balance under natural conditions. For the actual production of livestock, the ideal sex ratio is of great significance. Production enterprises using lactation and egg production traits hope to obtain more female individuals, while those with meat production and hair production traits have more obvious advantages in male individuals. Humans have always been interested in controlling the sex of animal offspring. At present, gender control is mainly carried out from two aspects: pre-fertilization and post-fertilization. The former uses sperm separation and then fertilizes, so that the sex ...

Examples

Embodiment 1

[0055] 1. Construct the RNAi interference vector of zfx gene:

[0056] The zfx gene sequence used comes from the NCBI gene bank, and 10 siRNA fragments were screened out according to the RNAi design principles and the comparative analysis results on NCBI, as shown in Table 1, which are used for siRNA zfx cDNA template. Add EcorI and BamI restriction sites at the 5' and 3' of each siRNA fragment, respectively, as shown in Table 2, and then send the siRNA fragment to be synthesized by a biological company. The positive and antisense strands were annealed into double strands and connected to the RNAi-Ready pSIREN-RetroQ-ZsGreen Vector (Clontech, USA) carrier, and then transformed into E. coli DH5α competent cells (Tiangen, China) for amplification. The extracted plasmid was stored at -20°C.

[0057] 2. In vitro interference test of zfx gene:

[0058] Four 28-day-old healthy male Kunming mice were selected, killed by cervical dislocation, and their abdomens were disinfected wi...

technical field

[0001] The present invention relates to the field of molecular genetics, in particular to the ability to silence zfx RNA interference fragments of genes and their applications. Background technique

[0002] Under natural conditions, the sex ratio of almost all mammals is close to 1:1, thus achieving uninterrupted and stable reproduction of offspring and achieving ecological balance under natural conditions. For the actual production of livestock, the ideal sex ratio is of great significance. Production enterprises using lactation and egg production traits hope to obtain more female individuals, while those with meat production and hair production traits have more obvious advantages in male individuals. Humans have always been interested in controlling the sex of animal offspring. At present, gender control is mainly carried out from two aspects: pre-fertilization and post-fertilization. The former uses sperm separation and then fertilizes, so that the sex ...

Examples

Embodiment 1

[0055] 1. Construct the RNAi interference vector of zfx gene:

[0056] The zfx gene sequence used comes from the NCBI gene bank, and 10 siRNA fragments were screened out according to the RNAi design principles and the comparative analysis results on NCBI, as shown in Table 1, which are used for siRNA zfx cDNA template. Add EcorI and BamI restriction sites at the 5' and 3' of each siRNA fragment, respectively, as shown in Table 2, and then send the siRNA fragment to be synthesized by a biological company. The positive and antisense strands were annealed into double strands and connected to the RNAi-Ready pSIREN-RetroQ-ZsGreen Vector (Clontech, USA) carrier, and then transformed into E. coli DH5α competent cells (Tiangen, China) for amplification. The extracted plasmid was stored at -20°C.

[0057] 2. In vitro interference test of zfx gene:

[0058] Four 28-day-old healthy male Kunming mice were selected, killed by cervical dislocation, and their abdomens were disinfected wi...

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