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Journal of Integrative Agriculture
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Inhibition of KU70 and KU80 by CRISPR interference, not NgAgo interference, increases the efficiency of homologous recombination in pig fetal fibroblasts
LI Guo-ling, QUAN Rong, WANG Hao-qiang, RUAN Xiao-fang, MO Jian-xin, ZHONG Cui-li, YANG Huaqiang, LI Zi-cong, GU Ting, LIU De-wu, WU Zhen-fang, CAI Geng-yuan, ZHANG Xian-wei
2019, 18 (
2
): 438-448. DOI:
10.1016/S2095-3119(18)62150-1
Abstract
(
276
)
PDF
(765KB)(
580
)
Non-homologous end-joining (NHEJ) is a predominant pathway for the repair of DNA double-strand breaks (DSB). It inhibits the efficiency of homologous recombination (HR) by competing for DSB targets. To improve the efficiency of HR, multiple CRISPR interference (CRISPRi) and
Natronobacterium gregoryi
Argonaute (NgAgo) interference (NgAgoi) systems have been designed for the knockdown of NHEJ key molecules,
KU70
,
KU80
, polynucleotide kinase/phosphatase (PNKP), DNA ligase IV (
LIG4
), and
NHEJ1
. Suppression of
KU70
and
KU80
by CRISPRi dramatically promoted (
P
<0.05) the efficiency of HR to 1.85- and 1.58-fold, respectively, whereas knockdown of PNKP, LIG4, and NHEJ1 repair factors did not significantly increase (
P
>0.05) HR efficiency. Interestingly, although the NgAgoi system significantly suppressed (
P
<0.05)
KU70
,
KU80
,
PNKP
,
LIG4
, and
NHEJ1
expression, it did not improve (P>0.05) HR efficiency in primary fetal fibroblasts. Our result showed that both NgAgo and catalytically inactive Cas9 (dCas9) could interfere with the expression of target genes, but the downstream factors appear to be more active following CRISPR-mediated interference than that of NgAgo.
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Genetic parameters and trends for production and reproduction traits of a Landrace herd in China
ZHANG Zhe, ZHANG Hao, PAN Rong-yang, WU Long, LI Ya-lan, CHEN Zan-mou, CAI Geng-yuan, LI Jia-qi, WU Zhen-fang
2016, 15 (
05
): 1069-1075. DOI:
10.1016/S2095-3119(15)61105-4
Abstract
(
1677
)
PDF in ScienceDirect
The objectives of this study were to estimate the genetic parameters and the breeding progress in a Landrace herd in China, and to predict the potential benefits by applying new breeding technology. Hereby, the performance records from a Landrace swine herd in China, composing over 33 000 pigs born between 2001 and 2013, were collected on six economically important traits, i.e., average daily gain between 30–100 kg (ADG), adjusted backfat thickness at 100 kg (BF), adjusted days to 30 kg (D30), adjusted days to 100 kg (D100), number born alive (NBA), and total number born (TNB). The genetic parameters were estimated by restricted maximum likelihood via DMU, and realized genetic trends were analyzed. Based on the real population structure and genetic parameters obtained from this herd, the potential genetic trends by applying genomic selection (GS) were predicted via a computer simulation study. Results showed that the heritability estimates in this Landrace herd were 0.55 (0.02), 0.42 (0.01), and 0.12 (0.01), for BF, D100, and TNB, respectively. Favorable genetic trends were obtained for D100, BF, and TNB due to direct selection, for ADG and NBA due to indirect selection. Long-term selection against D100 did not improve D30, though they are highly genetically correlated (0.64). Appling GS in such a swine herd, the genetic gain can be increased by 25%, or even larger for traits with low heritability or individuals without phenotypes before selection. It can be concluded that conventional breeding strategy was effective in the herd studied, while applying GS is promising and hence the road ahead in swine breeding.
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